Cell culture models of oral mucosal barriers: A review with a focus on applications, culture conditions and barrier properties

Figures & data

Table 1. Human tumor-derived cell lines of oral mucosa.

Cell Line	Studies/Applications/Notes	References^1
BUCCAL CARCINOMA
TR146	epithelial barrier studies, model cell line for human buccal epithelium (i.a. drug delivery, trans–epithelial permeability) model cell line for OSCC (i.a. cytotoxicity studies), C. albicans infection studies, 3D organotypic culture (i.a. in commercially available systems of reconstituted oral mucosa, e.g. SkinEthic Laboratories)	^Citation62∏, Citation74#, Citation76, Citation78, Citation83, Citation94–Citation97 barrier/permeability studies see also Table 3 >50 publications (“TR146”)
SqCC/Y1	3D organotypic culture, epigenetic studies, model cell line for malignant buccal carcinoma (i.a. in comparison to normal or immortalized keratinocytes such as SVpgC2a, see Table 3)	^Citation98∏, Citation99–Citation102, Citation103# >40 publications (“SqCC/Y1”)
HO-1-N-1 (Nakata-1; JCRB0831)	wound healing assays/migration, metastasis studies, investigation of retinoic acid (receptors) in OSCC	^Citation104∏, Citation105–Citation107#, Citation108,Citation109 >10 publications (“HO-1-N-1”)
H413 (06092007 SIGMA)	P. gingivalis infection studies, epithelial barrier studies, epigenetic studies, TGFβ studies	^Citation110∏, Citation111–Citation114#, Citation115 barrier/permeability studies see also Table 3 >10 publications (“H413 oral”)
YD-9	1 out of 8 YD oral cancer cell lines (see ^Citation116), chemoresistant SCC model cell line, sensitivity to anticancer drugs, cancer-related over-expression studies	^Citation116∏, Citation117–Citation120# >10 publications (“YD-9” or “YD9)
OC3	areca nut-chewing related buccal carcinoma cell line, cytotoxicity studies/tumor suppression	^Citation121Л, Citation122,Citation123#, Citation124,Citation125 >10 publications (“OC3 oral”)
TW2.6	areca nut-chewing and tobacco smoke-related buccal carcinoma cell line, tumor aggressiveness/migration/invasion studies, epigenetic studies	^Citation126∏, Citation127#, Citation128–Citation130 >10 publications (“TW2.6”)
TONGUE CARCINOMA
CAL27	OSCC model cell line, cancer invasion/migration/metastasis/drug sensitivity studies, …	^Citation131Л; recent publications: ^Citation132#, Citation133,Citation134 >100 publications (“Cal27”)
CAL33	OSCC model cell line, cancer invasion/migration/metastasis/drug sensitivity studies, …	^Citation131Л, recent publications: ^Citation135–Citation137 >40 publications (“Cal33”)
WSU-HN6	cancer cell migration/motility/overexpression studies, drug sensitivity	^Citation138Л, Citation139–Citation141 >10 publications (“WSU-HN6”)
H357 (06092004 SIGMA)	OSCC model cell line for several cancer–related studies: drug sensitivity, cell migration, invasion, aggressiveness, …	^Citation110Л; recent publications: ^Citation142–Citation144 >40 publications (“H357 cancer”)
SCC25 (ATCC® CRL-1628™)	1 out of 6 “SCC” cell lines, weakly invasive (see ^Citation145), OSCC model cell line for several cancer–related studies: drug sensitivity, signaling, migration, invasion, …	^Citation145Л; recent publications: ^Citation133,Citation146,Citation147 ~>300 publications (“SCC25” or “SCC-25”)
D20	moderate dysplastic cell line, 1 out of 16 dysplasias of the oral cavity (see ^Citation148), 3D organotypic culture of oral dysplasia	^Citation148Л, Citation149–Citation152# > 5 publications (“D20 dysplasia”)
DOK “dysplastic oral keratinocyte“	early and mild/moderate dysplastic cell line, tumor progression/migration studies, 3D organotypic culture of oral dysplasia, analysis of stemcell–associated markers	^Citation153Л, Citation150,Citation154–Citation157# >15 publications (“DOK dysplastic”)
OSCC-BD “oral squamous cell carcinoma–B(a)P/DMBA”	established from DOK by B(a)P/DMBA mixture (key carcinogens of tobacco smoke), identification of malignant transformation–related cancer proteins	^Citation158∏ 1 publication (“OSCC-BD”)
HSC-3 (JCRB0623)	OSCC model cell line for several cancer–related studies: drug sensitivity, cell migration, invasion, aggressiveness, cell signaling, stem cell markers …	^Citation159Л, recent publications: ^Citation160–Citation162 ~>300 publications (“HSC-3 oral” or “HSC3 oral”)
HSC-4 (JCRB0624)	OSCC model cell line for several cancer–related studies: drug sensitivity, cell migration, invasion, aggressiveness, cell signaling, stem cell markers …	^Citation163Л, recent publications: ^Citation160,Citation164,Citation165 ~>150 publications (“HSC-4 oral” or “HSC4 oral”)
OSC-19	low-grade invasive cell model of epithelial phenotype, metastatic-lesion derived, metastasis studies, epigenetic studies, cytotoxicity studies, drug sensitivity studies, …	^Citation166Л, Citation167–Citation170# >50 publications (“OSC-19 oral” or “OSC19 oral”)
OSC-20	low-grade invasive cell model of epithelial phenotype, metastatic-lesion derived, metastasis studies, cancer stem cell studies, …	^Citation171Л, Citation168,Citation170#,Citation172 >20 publications (“OSC-20 oral” or “OSC20 oral”)
SCCKN	mouse xenograft cell model, cytotoxicity/drug sensitivity studies, …	^Citation173Л, Citation174–Citation176# >20 publications (“SCCKN”)
PE⁄CA-PJ15	cytotoxicity/drug sensitivity studies, 3D organotypic culture, …	^Citation177Л, Citation156,Citation178–Citation180# >15 publications (“SCCKN”)
YD-8	1 out of 8 YD oral cancer cell lines (see ^Citation116), SCC, cytotoxicity/drug sensitivity studies, …	^Citation116Л, Citation181–Citation183# >5 publications (“YD-8” or “YD8)
YD-10B	1 out of 8 YD oral cancer cell lines (see ^Citation116), SCC, p53-mutant OSCC cell line, cytotoxicity/drug sensitivity studies, …	^Citation116Л, Citation182, Citation183#, Citation184, Citation185 >20 publications (“YD-10B” or “YD10B)
YD-15	1 out of 8 YD oral cancer cell lines (see ^Citation116), mucoepidermoid carcinoma, cytotoxicity/drug sensitivity studies, …	^Citation116Л, Citation186–Citation188# >10 publications (“YD-15” or “YD15”)
SCC-4 (ATCC® CRL-1624)	OSCC model cell line, metastasis/migration studies, cytotoxicity/drug sensitivity studies, …	^Citation189Л, recent publications: ^Citation190, Citation191# >100 publications (“SCC-4 oral” or “SCC4 oral”)
SCC-9 (ATCC® CRL-1629)	OSCC model cell line, metastasis/migration/invasion studies, overexpression studies, cytotoxicity/drug sensitivity studies, …	^Citation145Л, recent publications: ^Citation192,Citation193 >100 publications (“SCC-9 oral” or “SCC9 oral”)
SCC-15 (ATCC® CRL-1623)	OSCC model cell line, metastasis/migration/invasion studies, overexpression studies, cytotoxicity/drug sensitivity studies, …	^Citation189Л, recent publications: ^Citation194,Citation195# >100 publications (“SCC-15 oral” or “SCC15 oral”)
UMSCC2	metastatic SCC cell line, cytotoxicity/drug sensitivity studies, mouse xenograft cell model	^Citation196–Citation198# >10 publications (“UMSCC2”)
SAS	cytotoxicity/drug sensitivity studies, cancer stem cell properties, migration/invasion studies, …	recent publications: ^Citation191Л, Citation199,Citation200 ~>200 publications (“SAS oral carcinoma”)
GINGIVAL CARCINOMA
Ca9-22	OSCC model cell line, cytotoxicity/drug sensitivity studies, epigenetic studies, cancer-related expression studies, …	^Citation201Л, recent publications: ^Citation200,Citation202 >200 publications (“Ca9-22”)
MSCC-1	metastastic-lesion derived, metastasis/migration/invasion studies,	^Citation203Л, Citation204 2 publications (“MSCC-1”)
KOSC-3	invasion studies, mutational analysis	^Citation205Л, Citation206–Citation208# >5 publications (“KOSC-3” or “KOSC3”)
YD-38	1 out of 8 YD oral cancer cell lines (see ^Citation116), cytotoxicity/drug sensitivity studies, studies on tumor-associated macrophages	^Citation116Л, Citation117,Citation209,Citation210# >10 publications (“YD-38” or “YD38”)
OEC-M1	Metastasis/migration/invasion studies, cytotoxicity/drug sensitivity studies, cancer-, metastasis- related (over-)expression studies	^Citation211Л, Citation213–Citation215# >40 publications (“OEC-M1 oral” or “OECM1 oral”)
CARCINOMA OF THE FLOOR OF THE MOUTH
KOSC-2	invasion studies, mutational analysis	^Citation205Л, Citation206,Citation207,Citation215# >5 publications (“KOSC-2” or “KOSC2”)
POE-9n	severe dysplastic oral keratinocytes, not immortal but extended culture lifespan, 3D organotypic culture/tissue engineering,	^Citation216Л, Citation150,Citation156,Citation217# >5 publications (“POE9n” or “POE-9n”)
HO-1-U-1 (Ueda 1; JCRB0828)	metastasis/invasion studies, epithelial barrier studies	^Citation90Л, Citation91,Citation93,Citation165#, Citation218 >15 publications (“HO-1-U-1” or “HO1U1”)
H376 (06092005 SIGMA)	nanoparticle transport studies, cancer-related expression studies	^Citation110Л, Citation219–Citation221# >10 publications (“H376 oral”)
H314 (06092003 SIGMA)	cancer-related expression studies, TGFβ–related studies	^Citation110Л, Citation222–Citation224# >5 publications (“H314 oral”)
OTHERS (or not specified)
KB (ATCC CCL-17) epidermoid carcinoma ! cross - contaminated with HeLa cells! continued misrepresentation of KB cells as oral cancerphenotype (see ^Citation225)	cancer-related (over-)expression studies, cytotoxicity/drug sensitivity studies, migration/metastasis/invasion studies …	^Citation226Л, recent publications: ^Citation120,Citation200,Citation227 >300 publications (“KB epidermoid carcinoma oral”)
H400 (06092006 Sigma) alveolar process squamous cell carcinoma	cytotoxicity/drug sensitivity studies, 3D organotypic culture	^Citation110Л, Citation228–Citation230# >10 publications (“H400 oral”)
MEMO oral mucosal melanoma cell line	established from unusual case of oral mucosal melanoma on hard palate and maxillary alveolar ridge	^Citation231∏ 1 publication, i.e. establishment (“MEMO melanoma”)
1483/SCC1483 carcinoma of retromolar trigone region of the oropharynx some stocks genetically identical to UM-SCC1 (see ^Citation233)	cancer-related expression studies, cytotoxicity/drug sensitivity studies	^Citation233Л, Citation234–Citation236 >10 publications (“1483 retromolar”)
FaDu hypopharyngeal squamous cell carcinoma	migration/invasion studies, cytotoxicity/drug sensitivity studies	^Citation237∏; recent publications: ^Citation238–Citation240 >80 publications (“FaDu oral”)
BHY squamous cell carcinoma of lower alveolus	cytotoxicity/drug sensitivity studies, cancer-related expression studies	^Citation241Л, Citation242–Citation244# >20 publications (“BHY oral”)
HSC-2 (JCRB0622) SCC of oral cavity	OSCC model cell line, cytotoxicity/drug sensitivity studies, cancer-related (over-) expression studies, metastasis/migration/invasion studies, …	^Citation163Л, recent publications: ^Citation165,Citation199,Citation245# >200 publications (“HSC-2 oral” or “HSC2 oral”)
HOC313 OSCC involving the mandibular gingiva and oral floor	high-grade invasive cell model of mesenchymal phenotype, metastatic-lesion derived, cancer progression/metastasis/EMT studies	^Citation246Л, Citation170#,Citation247, Citation248# >20 publications (“HOC313”)
SCC Cell Line Collections
PCI cell lines	21 HNSCC cell lines	^Citation249Л
JHU cell lines (“John Hopkins University”)	5 SCC cell lines of oral cavity: JHU-011 (larynx), −012 (oral cavity), −013 (oral cavity), −019 (base of tongue), −028 (unknown)	^Citation250
73 UM-SCC (“University of Michigan Squamous Cell Carcinoma”) cell lines	73 SCC cell lines of oral cavity	genotyping for all cell lines see ^Citation251

# … latest publication found using respective cell line

∏… publication describing establishment of respective cell line

1) References do not represent a complete list of publications, but list exemplary and/or latest studies. Approximate number of publications should give an impression of how often respective cell lines have been used previously. Search terms are given in parentheses.

abbreviations: HNSCC, head and neck squamous cell carcinoma; OSCC, oral squamous cell carcinoma; SCC, squamous cell carcinoma;

Table 2. Immortalized Cell Lines of Oral Mucosa.

Cell Line	Immortalization	Studies/Applications	References^Citation1
BUCCAL MUCOSA
SVpgC2a human	SV40T	3D organotypic culture, model cell line for immortalized/transformed buccal keratinocytes, head and neck cancer biomarker study	^Citation28∏, Citation99, Citation253, Citation100, Citation282#
NOM9-CT human	Cdk4 + hTERT	effect of curcumin treatment	^Citation283∏#⌂
IMOK mouse buccal and palatal mucosa	spontaneous	first cell line to recapitulate mouse oral epithelium differentiation	^Citation284)∏, Citation285# ⌂
foec (clonal cell lines 1–8) mouse	p53–deficient mouse	foec-6, −8: 3D organotypic culture, foec-2, −3, -5, −6, −7, and −8: preparation of bioengineered tooth germ	^Citation286∏# ⌂
GINGIVA
HMK human	spontaneous	3D organotypic culture, HPV/HSV infection studies	^Citation287∏, Citation288, Citation289, Citation290, Citation291#
IHGK human	HPV16 E6/E7	normal keratinocytes control cell line for cancer studies, 3D organotypic culture, nicotine studies	^Citation292∏ Citation293, Citation294, Citation295, Citation296#
MOE (1a and 1b) human	MOE1a: mutant Cdk4, Cyclin D1, hTERT, dominant–negative p53mutant MOE1b: mutant Cdk4, Cyclin D1, hTERT	immortalization process without viral oncogenes	^Citation297∏#⌂
GE1 mouse	SV40T	epithelial barrier studies, gingival gene expression studies	^Citation257∏, Citation260, Citation258, Citation259, Citation298, Citation299)#
TIGK human	bmi1/hTERT	bacterial infection studies (mainly P.gingivalis)	^Citation300∏, Citation301, Citation302, Citation303, Citation304#
HGEK-16 human	HPV16 E6/E7	3D organotypic culture	^Citation305∏#⌂
NDUSD	SV40T	cytological and cytogenetic characterization	^Citation306∏#⌂
human
OKG4/bmi1/TERT	bmi1/TERT	3D organotypic culture	^Citation216∏, Citation296#
human
MCNR 3B11 =  Gie-No3B11 human	HPV16 E6/E7	epithelial barrier studies, bacterial infection studies (P.gingivalis)	^Citation254∏, Citation255, Citation256#⌂
OBA-9 human	SV40T	bacterial infection studies	^Citation307∏, Citation308, Citation309, Citation310, Citation311#
NOK-SI human	spontaneous	epithelial stem cell functions, wound healing/migration, normal keratinocyte control cell line in cancer and overexpression studies, epigenetics …	^Citation312∏, Citation313, Citation314, Citation315, Citation316#
FLOOR OF THE MOUTH
OKF-6/TERT2 human	hTERT	3D organotypic culture (oral mucosa equivalents, periodontal biofilm model, bone–oral mucosa model), stemcell–associated marker expression, infection studies (bacteria, C.albicans), toxicological studies …	^Citation216∏, Citation281, Citation317, Citation318, Citation319, Citation320, Citation156, Citation321, Citation322, Citation323#
LABIAL VESTIBULE
GMSM-K human	SV40T	bacterial infection studies, model cell line for normal keratinocytes	^Citation324∏, Citation325, Citation326, Citation327#⌂
TONGUE
ROE2 rat	SV40T	toxicological studies of fluoride	^Citation328∏, Citation329#⌂

# … latest publication found using respective cell line

⌂ … complete list of publications using respective cell line is given

∏… publication describing establishment of respective cell line

1 references do not represent a complete list of publications, but list exemplary and/or latest studies; unless marked with ⌂

abbreviations: Cdk4, cyclin – dependent kinase 4; HPV16 E6/E7, human papillomavirus 16 E6/E7 proteins; hTERT, human telomerase reverse transcriptase; SV40T, simian virus 40 T antigen

Table 3. Oral Mucosa Cell Models used for In vitro Barrier Studies.

Cell Line	Cell Culture Medium	Model Set – Up (permeability assays)	Studied Barrier Properties	References
tumor-derived
TR146 human buccal carcinoma	DMEM + 10% FCS + 50 µg/ml gentamicin + 0.2 µg/ml p-hydroxybenzoic acid n-butyl ester DMEM + 10% FCS + 3.7 mg/ml NaHCO3 + 50 µg/ml gentamicin + 0.2 µg/ml p-hydroxybenzoic acid n-butyl DMEM + 10% FCS + 100 IU/ml penicillin + 100 µg/ml streptomycin DMEM + 10% FBS + 200 µM L-glutamine + 100 IU/ml penicillin + 100 µg/ml streptomycin	polyethylene terephthalate filters, Becton Dickinson, 1.6x10^6 pores/cm^2, 0.45 µm pore size, 4.6 cm^2 growth area, 2.4x10^4 cells/cm^2; media change: 3x/week 12-well polyethylene terephthalate filters, Becton Dickinson, 0.4 µm pore size, 0.9 cm^2 growth area, 2.1x10^4 cells/filter polyethylene terephthalate filters, Becton Dickinson, 1.6x10^6 pores/cm^2, 0.40 µm pore size, 4.2 cm^2 growth area, 2.4x10^4 cells/cm^2 polyethylene terephthalate filters, Becton Dickinson, 0.40 µm pore size, 4.2 cm^2 growth area, 2.4x10^4 cells/cm^2 12 well polyethylene terephthalate filters, Becton Dickinson, 0.4 µm pore size, 0.9 cm^2 growth area, 2.2x10^4 cells/cm^2	-TEER^Δ: max. TEER at d30: 55–120 Ωcm^2 (inter-passage variation) -tracer flux: mannitol (hydrophilic), min. Papp at d30: ~ 4x10^−6 cm/s; testosterone (lipophilic), Papp independent of d: ~ 2x10^−5 cm/s -TEM: intermediate filaments, microvilli-like processes, no TJs, membrane coating granules, absence of complete keratinization -TEER^Δ: 339 ± 89 Ωcm^2 (d28-d30) -tracer flux: ^3H mannitol, FITC-dextran (4 kDa, 10 kDa, 20 kDa); enhancement of permeability by chitosan glutamate; e.g. 4 kDa FITC-dextran: Pc = 7.85 ± 2.22 (x10^7) cm/s increased by 20 µg/ml chitosan to Pc = 22.52 ± 6.60 (x10^7) cm/s; 10 kDa FITC-dextran: Pc = 2.95 ± 1.18 (x10^7) cm/s increased by 20 µg/ml chitosan to Pc = 5.32 ± 0.44 (x10^7) cm/s (more values see Table II in ^284) -TEER^Δ: max. TEER at d23, submerged culture: 102 ± 5 Ωcm^2 -tracer flux: ^14C mannitol, min. Papp at d23, submerged culture: 4.08 ± 0.15x10^−6 cm/s -TEER^Δ: 154 ± 13 Ωcm^2 (~ d21), decreased by 10 mM GC treatment: 92 ± 49 Ωcm^2 -tracer flux: FITC-dextran (4–40 kDa): decreasing Papp with increasing Mw 0.65 ± 0.055x10^−8 cm/s to 44 ± 7.5x10^−8 cm/s; -TEER^Δ: 151 ± 38 Ωcm^2 (d27-d29), decreased by chitosan treatment (up to 20 mg/ml) -tracer flux: ^14C mannitol, Papp: 3.79 ± 0.68x10^−6 cm/s, increased by chitosan treatment (up to 20 mg/ml) test of different polymers (chitosan, LM-pectin, HM-pectin, AM-pectin, Eudragit, p(NIPAAM-co-MAA), HM-HEC, HM-EHEC) on cell permeability: -TEER^Δ: significant decrease only by chitosan (d27-d29), but reduced cell viability -tracer flux: ^14C mannitol: significant increase of Papp only by chitosan (d27-d29), but reduced cell viability	^Citation63 ^Citation73 ^Citation64 ^Citation65 ^Citation72 ^Citation74
HO-1-u-1 (Ueda-1) human SCC of the floor of the mouth	DMEM + 10% FCS + 90 IU/penicillin + 90 µg/ml streptomycin Ham’s F-12 + 10% FCS + 2 mM glutamine + 100 IU penicillin/streptomycin + 10 μg gentamicin DMEM-F12 (1:1) + 10% FCS + 50 µg/ml gentamicin + 100 IU/ml penicillin + 100 µg/ml streptomycin	12 well PC filters, Corning Costar, 3.0 µm pore size, 1.131 cm^2 growth area, 2.4x10^4 cells/cm^2 polyethylene terephthalate filters, Becton Dickinson, 0.4 µm pore size, 0.9 cm^2 growth area 12 well PET filters, Appletonwoods, 2.4x10^4 cells/cm^2; media change: every 2–3 days 6 well polyethylene terephthalate inserts, Becton Dickinson, 0.45 µm pore size; media change: 3-4x/week 6 well polyethylene terephthalate inserts, Becton Dickinson, 0.45 µm pore size, 1.5x10^5 cells/well; media change: 3-4x/week	-TEER^Δ: 50.02 ± 2.87 Ωcm^2 (d27-d28) -transport studies of polystyrene nanoparticles using an advanced TR146 model including a mucus layer -TEER^Δ: 36.4 ± 7.6 Ωcm^2 (d28-d30); 50% decrease by ~ 12 mM GC treatment -tracer flux: ^3H mannitol, Papp: 3.3 ± 0.4x10^−6 cm/s, increased to 6.4 ± 0.1x10^−6 cm/s by ~ 12 mM GC treatment -TEER^Δ: ~ 200 Ωcm^2 (d30); no significant changes by lysine (10 µg/ml), histidine (10 µg/ml), glutamic acid (100 µg/ml, 200 µg/ml) and aspartic acid (200 µg/ml) treatment -transport studies: insulin, max. permeation: 4.07 ± 0.82% (4h); significant enhancement of insulin permeation by lysine (10 µg/ml), histidine (10 µg/ml), glutamic acid (100 µg/ml, 200 µg/ml), aspartic acid (200 µg/ml) treatment -TEER^Δ: time-course (d8–d30): continuous increase from 71.4 ± 9.8 Ωcm^2 to a maximum of 326.3 ± 41.3 Ωcm^2 (d17), then slowly decrease to 219.1 ± 28.8 Ωcm^2 (d30) -tracer flux: ^14C mannitol (hydrophilic), Papp: decrease from 3.64 ± 0.28 × 10^−6 cm/s (d15) to 2.04 ± 0.17 × 10^−6 cm/s (d26), then slightly increase to 2.42 ± 0.29 × 10^−6 cm/s (d29); ^3H testosterone (lipophilic), Papp no significant change until d29, ~ 5x10^−6 cm/s; 8 different β-blockers (compounds involving passive diffusion), Papp: range from 2.89 ± 0.17 × 10^−6 cm/s (atenolol) to 6.37 ± 0.37 × 10^−6 cm/s (alprenolol) (d2) -TEM: polarized structure, 2–3 cell layers, short microvilli-like processes, desmosomes, no TJs (d23) -TEER^Δ: no significant differences at various pH (3.5–10), e.g. 175.4 ± 2.5 Ωcm^2 at pH 3.5, 167.5 ± 9.2 Ωcm^2 at pH 10 -tracer flux: ^14C mannitol, ^3H testosterone (neutral markers), Papp: independent of solution pH, e.g. ^14C mannitol with 1.92 ± 0.18x10^−6 cm/s at pH 3.5, 1.63 ± 0.14x10^−6 cm/s at pH 10; β-blockers [aterolol (hydrophilic), metoprolol (lipophilic), propranolol (lipophilic)], several Papp at different osmolarities and GDC concentrations	^Citation87 ^Citation330 ^Citation75 ^Citation91 ^Citation93
H413 human buccal carcinoma	MEM + 5% FCS (low calcium medium, 0.2 mM calcium contributed from FCS)	24 mm Transwell PET filters, 0.4 µm pore size, Corning, 2 × 10^5 cells/cm^2	-tracer flux: monolayers impermeable to tetramethylrhodamine dextran (2 MDa), permeable to Dextran Alexa Fluor 647 (10 kDa); + anti-CD24: reduced permeability -TJ/AJ expression: PCR: + anti-CD24: upregulation: ZO-1, ZO-2, Occl WB: + anti-CD24: upregulation: ZO-1, ZO-2, Occl -tracer flux: Dextran Alexa Fluor 647 (10 kDa); + 0.25 ng/ml TGFβ3: decreased permeability (3h); increased permeability from 4h treatment -TJ/AJ expression: gene expression/protein analysis of 28 TJ components in response to 0.25 ng/ml TGFβ3: PCR: 1) marked up-regulation: Cldn4 2) moderate/low up-regulation: Cldn20, Cldn18 A2.1, Cldn 14, Cldn 15, Cldn 10a, ZO-2;/JAM-B, Cldn11 3) marked down-regulation: Cldn1 4) moderate/low down-regulation: Cldn2, Cldn12, Cldn8, Cldn6, JAM-C 5) no regulation: JAM-A, ZO-1, ZO-3, Occ, Cldn7, Cldn9, Cldn16, Cldn17, Cldn22 6) no expression: Cldn10b, Cldn3, Cldn5, Cldn19, Cldn23, Cldn18A1.1	^Citation331 ^Citation113
immortalized MCNr3B11 (Gie-No3B11) human gingival keratinocytes, HPV–16 E6/E7 immortalization	1. DMEM–Ham’s F12 (4:1) induction of differentiation: 2. DMEM–Ham’s F12 (4:1) + 10% FCS + 1.8 mM calcium + 10 mM HEPES buffer + 200 mM L-glutamine 1. DMEM–Ham’s F12 (4:1) + HEPES induction of differentiation: 2. DMEM–Ham’s F12 (4:1) + 10% FCS + 1.8 mM calcium	24 well Transwell-Col® filter inserts, Corning Costar, 3.25x10^5 cells/insert, 24 well ThinCert^TM filter inserts, Greiner Bio One, 3.25x10^5 cells/insert	WB: 1) significant decrease: Cldn2, Cldn4, Cldn10 2) significant increase: Cldn20, Occ- TEER^Δ: ~ 160 Ωcm^2 (d1); stable until d3, decrease from d4 -TJ/AJ expression: IF: Cldn1^pos (cellular walls), Cldn2^pos, Occ^pos -TEER^Δ: ~ 110 Ωcm^2 (d1) + 15 µM RA: 127 Ωcm^2 (after 1h), 150 Ωcm^2 (after 4h), 189 Ωcm^2 (after 6h) -TJ/AJ expression: WB: Cldn1, Cldn2, Cldn4, Occ, ZO-1 + 15 µM RA: increased expression of Cldn4, Occ; decreased expression of ZO-1; no effect on expression of Cldn1, Cldn2;	^Citation254, Citation255 ^Citation256
GE-1 mouse gingival keratinocytes, temperature -sensitive SV40T immortalization	SFM101 + 1% FCS + 10 ng/ml EGF	-	.-TJ/AJ expression: IF: Cldn1^pos (cellular walls), Cldn4^pos (cellular walls of medial layers), Cldn5^neg, ZO–1^mod + RA: Cldn1^pos (cytoplasm), Cldn4^pos (cellular walls), Cldn5^pos, ZO–1^pos PCR: Cldn1, Cldn4, Cldn5, Occ, ZO-1 + RA: increased expression of Cldn4, Occ, ZO–1 (n.s.); decreased expression of Cldn1, Cldn5; -gap junction expression: IF: Cx26^pos, Cx31.1^pos, Cx32^pos, Cx43^pos PCR: Cx26, Cx30.3, Cx31.1, Cx32, Cx43 + RA treatment: reduced expression of Cx31.1 and Cx32 -TEM: stratified epithelium, desmosomes, hemidesmosomes, anchoring tonofilaments; + RA: thinner cell layer, loss of tonofibrils, less and shorter desmosomes, complete loss of hemidesmosomes -anchoring junctional molecules expression: IF: desmosomes: K13^pos, K4^pos, desmoglein 1^pos + RA: All decreased PCR: desmoglein 1, desmocollin 1, K13, K14, BPAG1, BPAG2, plectin, laminin 5 + RA: decreased expression of desmoglein 1, desmocollin 1, K13, K14, BPAG1; increased expression of plectin; no changes for BPAG2 and laminin 5	^Citation258 ^Citation259 ^Citation260
primary human gingival keratinocytes	DMEM + 15% FCS KGM	24 well cellagen® filter inserts, Costar, 4 × 10^5 cells/insert 12 mm Transwell® PC filter inserts, Corning, 0.4 µm pore size, 5 × 10^4 cells/insert	-TEER^Δ: 374 Ωcm^2 (d5), decrease to 120 Ωcm^2 (d13) -TEM: tight junctions, some desmosomes, no gap junctions -TEER^Δ: ~ 100 Ωcm^2 (~ d3) -TJ/AJ expression: IF: poor detection of Cldn1 (data not shown), no formation of TJs PCR: increased expression of E-cad, β-cat and β-actin by bacterial toxin Cdt (in majority of examined donors)	^Citation265 ^Citation261
human gingival keratinocytes	KGM	0.40 µM polyethylenetetraphthalate track-etched membranes, BD Biosystems	-TEER^Δ: 176–208 Ωcm^2 (independent biopsie samples) -TJ/AJ expression: no expression of ZO proteins (data not shown)	^Citation294
human soft palate keratinocytes	CnT-30 corneal epithelium medium + 1mM CaCl2 after 1 day of submerged culture, 6 days airlift in: 1) CnT-30 corneal epithelium medium 2) CnT-30 corneal epithelium medium + 10 ng/ml EGF 3) CnT-30 corneal epithelium medium + conditioned medium from human foreskin fibroblasts 4) CnT-30 corneal epithelium medium + conditioned medium from human foreskin fibroblasts + 10 ng/ml EGF	mouse collagen IV–coated filter inserts, BD Bioscience, 1 µm pores; media change: every day or every 2–3 days	-TEER^Δ: medium 1) 2379 ± 1167 Ωcm^2 medium 2) 2205 ± 1200 Ωcm^2 medium 3) 2007 ± 1119 Ωcm^2 medium 4) 1610 ± 439 Ωcm^2 (all d13-d17) -TJ/AJ expression: indirect IF: ZO–1^pos, Occ^pos (medium 1)	^Citation267,332
human gingival keratinocytes (EpiGingival^TM 3D model, MatTek)	KGM + 0.05 mM CaCl2 + 200 mM L-glutamine	24 well plate format	-TJ/AJ expression: PCR: mRNA expression of 62 genes (TJs, AJs, gap junctions; see Table 1 of publication ^297)	^Citation266

d … days in culture

Δ … TEER readings from filters without cells were subtracted from values obtained with filters and seeded cells

abbreviations: AJ, adherens junction; AM – pectin, amidated pectin; β-cat, β-catenin; CK, cytokeratin; Cldn, claudin; DMEM, Dulbecco’s Modified Eagle Medium; E-cad, E – cadherin; EGF, epidermal growth factor; FCS, fetal calf serum; GC, sodium glycocholate; GDC, sodium glycodeoxycholate; HM-EHE, hydrophobically modified ethyl hydroxyethyl cellulose; HM-HEC, hydrophobically modified hydroxyethyl cellulose;HM-pectin, highmethoxylated pectin; hTERT, human telomerase reverse transcriptase; IF, immunofluorescence; JAM-A, junctional adhesion molecule A; LM-pectin, low-methoxylated pectin; MUC, mucin; Occ, occludin; p(NIPAAM-co-MAA, poly(N-isopropylacrylamide-co-methacrylic acid); P_app, apparent permeability coefficient; P_c, cellular permability coefficient; PC, polycarbonate; PET, polyester; SCC, squamous cell carcinoma; TEER, transepithelial electrical resistance; TEM, transmission electron microscopy; TJ, tight junction; WB, Western Blot; ZO, zonula occludens.

Table 4. SALIVARY GLAND EPITHELIAL CELL LINES, This table summarizes cell models that have been used for the study of epithelial barrier properties of salivary glands.

Cell Line	Source	Cell Culture Medium	Characterization (ductal/acinar)	Model Set-Up (permeability assays)	Studied Barrier Properties	References
tumor-derived
HSG^# human	neoplastic submandibular gland intercalated duct epithelium	DMEM/F-12 (1:1)^1 + 10% FCS + 100 U/ml penicillin + 100 µg/ml streptomycin + 2.5 µg/ml amphotericin B^2	n.a. -non-coated: IF (d5): CK^neg, VIM^mod(88%), αSMA^neg, αAMY^neg, AQP5^neg, MUC1^neg -MG-coated: IF (d5): CK^pos, VIM^mod(61%), αSMA^neg, αAMY^pos, AQP5^pos, MUC1^mod(30%)	24 mm^3 Transwell PC filter, Corning Costar, 1 × 10^6 cells/well 24 mm Transwell-Clear PET filter, non-coated or MG-coated (f.c. 2 mg/ml, BD Biosciences), 5 × 10^4 cells/cm^2	-TEER^Δ: Ø50 Ωcm^2 (time-course, d0-d5)^4 -TJ/AJ expression: IF: ZO-1^neg, Occ^neg, Cldn1^neg, Cldn2^neg, β-cat^pos -TEER^Δ: -non-coated: 2,1,3 Ωcm^2 (d1, d3, d7) -MG-coated: 401, 417, 332 Ωcm^2 (d1, d3, d7) -TJ/AJ expression: IF (d5): -non-coated: Cldn1^neg, Cldn2^neg, Cldn3^neg, Cldn4^neg, Occ^neg, JAM-A^neg, ZO-1^neg -MG-coated: Cldn1^pos, Cldn2^pos, Cldn3^pos, Cldn4^pos, Occ^pos, JAM-A^pos, ZO-1^pos PCR (d3): -non-coated: Cldn1, Cldn2 -MG-coated: Cldn1, Cldn2 -TEM: -non-coated: small secretory granules, no Golgi apparatus, no rER, no microvilli -MG-coated: secretory-like granules, Golgi saccules, rER, TJs, no basal lamina	^Citation346 ^Citation347
CSG 120/7 mouse	submandibular gland carcinoma	DMEM + 10% FCS + 200 U/ml penicillin + 200 U/ml streptomycin	n.a. n.a.	12 mm Transwell filters, 0.4 µm pore size, Costar 12 mm Transwell PC filters, 0.4 µm pore size, Millipore	-TEER^Δ: e.g. ~ 250 Ωcm^2 (d2), 2350 ± 290 Ωcm^2 (d14) (time-course, d0-d14, measured every 2 days) -TJ/AJ expression: IF (d7): Occ^pos, ZO-1^pos, ZO-2^pos, JAM-A^pos, E-cad^pos, β-cat^pos -tracer flux: 4 kDa FITC-dextran (0.62 ± 0.39 ng/ml, d14) -tracer flux: [^3H]inulin (100%^5)	^Citation357 ^Citation358
immortalized
HSDEC human	-submandibular gland ductal epithelium -hTERT immortalization	BEBM + BEGM Single-Quots (Lonza) + 100 U/ml penicillin + 100 µg/ml streptomycin + 2.5 µg/ml amphotericin B	IF^6: CK7^pos, VIM^neg	12 mm Transwell filters, 0.4 µm pore size, Corning	-TEER^Δ: ~ 1700 Ωcm^2 (d5); + 100 ng/ml TNFγ: ~ 4000 Ωcm^2; + 100 ng/ml TGFβ: ~ 100 Ωcm^2 -TJ/AJ expression: IF: Cldn4^pos, Cldn7^pos, Occ^pos, JAM-A^pos + 100 ng/ml TNFγ: increased Cldn7 expression (WB, PCR), strong expression of Occ (IF); + 100 ng/ml TGFβ: lower expression of Occ at cell borders (IF)	^7
SMIE rat	-submandibular gland -adenovirus 12S E1A gene product immortalization	DMEM + 10% FCS + 100 U/ml penicillin + 100 μg/ml streptomycin	n.a. n.a. n.a.	24 mm Transwell filters, 0.4 μm pore size, Corning Costar, 10^5 cells/cm^2 24 mm Transwell types I and III collagen-coated PC inserts, Corning, 0.4 µm pore size, 3 × 10^6 cells/well 24.5 mm Transwell types I and III collagen-coated PC filter, Costar, 3 × 10^6 cells/well	-TEER^Δ: Ø58 Ωcm^2 (time-course, d4-d7); overexpression of GFP-Cldn4: ~ 40 Ωcm^2 (3d after plating, i.e. 2d after transfection); ~ 80 Ωcm^2 (4d after plating, i.e. 3 days after transfection) -TJ/AJ expression: IF (d4): ZO-1^pos, Occ^pos, Cldn3^pos, Cldn4^neg WB (d n.i.): E-cad, Occ, Cldn3, Cldn2^n.d., Cldn4^n.d., Cldn6-8^n.d., Cldn10^n.d., Cldn12^n.d. PCR (d n.i.): E-cad, Occ, Cldn3^7, Cldn12, Cldn1^n.d., Cldn2^n.d., Cldn4^n.d.- Cldn^11, Cldn14^n.d.–Cldn20^n.d., Cldn22^n.d., Cldn23^n.d. -tracer flux^8: 4 kDa FITC-dextran (~ 18% paracellular flux of input), 70 kDa FITC-dextran (~ 8% paracellular flux of input); overexpression of GFP-Cldn4: decreased flux of 4 kDa and 70 kDa FITC-dextran (4d after plating, i.e. 3 days after transfection) -TEER^Δ: ~ 40 Ωcm^2 (d10), serum-free: < 10 Ωcm^2 (d10) -TJ/AJ expression: IF (d10): ZO-1^pos, Occ^pos WB (d10): Cldn3, Occ, Cldn4^n.d. -tracer flux: 4 kDa FITC-dextran (~ 15% paracellular flux of empty filter control; serum-free: ~ 75% paracellular flux of control)^9 -TEER^Δ: ~ 30 Ωcm^2 (d n.i.) -TJ/AJ expression: IF (d7): ZO-1^pos	^Citation340 ^Citation359 ^Citation360
SMG-C6 rat	-submandibular gland acinar cells -replication defective SV40 immortalization	DMEM/F-12 (1:1) + 2.5% FCS + 5 μg/ml transferrin + 1.1 μmol/l hydrocortisone + 0.1 μmol/l retinoic acid + 2 nmol/l thyronine T3 + 5 μg/ml insulin + 80 ng/ml EGF + 50 μg/ml gentamicin sulfate + 5 mmol/l glutamine + 100 U/ml penicillin + 100 μg/ml streptomycin	WB (d n.i.): CK19^neg n.a.	12 mm Snapwell PC filters, 0.4 µm pore size, Costar, coated with 1.0 µg/cm^2 human collagen type I (Becton Dickinson), 2.5x10^5 cells/cm^2 6.5 mm Transwell filters, Corning, 0.4 μm pore size, 3.8x10^4 cells/cm^2	-TEER^Δ: 956 ± 84 Ωcm^2 (d n.i.) -TEER^Δ: 463.69 ± 13.35 Ωcm^2 (d6); + TNFα: concentration- + time-dependent decrease, e.g. to ~ 200 Ωcm^2 (50 ng/ml, 48h) -TJ/AJ expression: PCR ± WB (d n.i.): Cldn1, Cldn3, Cldn4, ZO-1, ZO-2, β-cat; + TNFα: concentration- + time-dependent disruption of Cldn3 distribution (WB, IF) -tracer flux:4 kDa + 40 kDa FITC-dextran (time course 0-48h, start d6)^10; + TNFα: 4 kDa and 40 kDa FITC-dextran flux significantly elevated (24h, 48h) ® overexpression, depletion or re-expression of Cldn3 altered the response of TNFα induced paracellular permeability and tight junction contents	^Citation345 ^Citation8
			n.a. n.a.	24 well Transwell chambers (0.33 cm^2 growth area), 0.4 µm pore size, Costar 24 well Transwell chambers (0.33 cm^2 growth area), PC filter, Costar, confluent density	-TEERΔ: ~ 600 Ωcm2 (d6/7)^11; no altered TEER by ZO-1, ZO-2, ZO-1/ZO-2 KD; significant drop in TEER by CAP (4-60min), but effect reversed in ZO-1, ZO-2, ZO-1/ZO-2 KD -TJ/AJ expression: PCR ± IF ± WB (d n.i.): ZO-1, ZO-2, ZO-3, Occ, β-cat -tracer flux: 4 kDa FITC-dextran^11; 4 kDa FITC-dextran flux significantly increased in ZO-1, ZO-2, ZO-1/ZO-2 KD, CAP significantly increased 4 kDa FITC-dextran flux, no effect in KD cells -TEER^Δ: 516.2 ± 98.62 Ωcm^2 (d5-d7); + CAP: concentration- + time-dependent decrease + Occ KD or ERK1/2 KD: significant decrease in TEER -TJ/AJ expression: IF (d n.i.): E-cad^pos, β-cat^pos, Cldn3^pos PCR (d n.i.): ZO-1, Occ, Cldn1, Cldn3, Cldn4, Cldn2^n.d., Cldn5^n.d. WB: Occ	^Citation361 ^Citation362
			n.a. n.a. n.a.	24 well Transwell chambers, PC filter (0.33 cm^2 growth area), 0.4 µm pore size, Costar 24 well Transwell chambers, PC filter, 0.33 cm^2 growth area, 0.4 µm pore size, Costar 24 well Transwell chambers, PC filter 0.33 cm^2 growth area, 0.4 µm pore size, Costar	-tracer flux:4 kDa FITC-dextran, trypan blue^12 + CAP: increased permeability for 4 kDa FITC-dextran and trypan blue -TEER^Δ: 632.8 ± 107.1 Ωcm^2 (d7); + Cch: concentration- + time-dependent decrease -TJ/AJ expression: WB (d n.i.): E-cad, Occ, ZO-1, Cldn1, Cldn4 IF (d n.i.): Cldn4^pos, Occ^pos, ZO-1^pos; + Cch: altered distribution of Cldn4 (IF) -tracer flux: 4 kDa + 40 kDa FITC-dextran^12; + Cch: concentration- + time-dependent increase of 4 kDa flux -TEER^Δ: 592.67 + 29.8 Ωcm^2 (d n.i.); + AICAR: decreased TEER values by ~ 46% (60min); Ca2+ free medium decreased TEER values by ~ 60% (60min); -TJ/AJ expression: WB ± IF (d n.i.): Occ, ZO-1, Cldn1, Cldn3, Cldn4 + AICAR: altered distribution of Cldn4 (IF) -tracer flux: 4 kDa FITC-dextran^12; + AICAR: increased permeability of 4 kDa FITC-dextran by 81.59% (60min) -TEER^Δ:635 ± 36.64 Ωcm^2 (d5/6/7); time-dependent decrease of TEER: + fAd (~ 60% decrease, 30min), + gAd (~ 60% decrease, 60min), + AICAR (~ 40% decrease, 60min)	^Citation342 ^Citation341 ^Citation363
Par-C10 rat	-parotid gland acinar cells -replication defective SV40 immortalization	DMEM-F12 (1:1) + 2.5% FCS + 0.1 μM retinoic acid + 80 ng/ml EGF + 2 nM thyronine T3 + 5 mM glutamine + 0.4 μg/ml hydrocortisone + 5 μg/ml insulin + 5 μg/ml transferrin + 5 ng/ml sodium selenite + 50 μg/ml gentamicin + 8.4 ng/ml cholera toxin	n.a. n.a.	n.i. 12 mm Falcon permeable supports, 0.4 µm pore size, Becton Dickinson, 5 × 10^5 cells/well	-TEM: secretory granules, TJ, intermediate junctions, desmosomes, microvilli -TEER^Δ: ~ 3500 Ωcm^2 (d4); + TNFα/INFγ: concentration- + time-dependent decrease, e.g. + 10 ng/ml TNFα: ~ 1200 Ωcm^2 (48h), + 10 ng/ml INFγ: ~ 2000 Ωcm^2 (48h), + 10 ng/ml TNFα/INFγ: ~ 400 Ωcm^2 (48h) -TJ/AJ expression: WB (d n.i.): Cldn1, Cldn3, Cldn4, Cldn10, Occ, ZO-1, JAM-A -tracer flux: 1.9 kDa/20Å microperoxidase MP-11, 44 kDa/50–60 Å horseradish peroxidase type VI, 80 kDa/120Å bovine milk lactoperoxidase^13; + TNFα, + INFγ, + TNFα/INFγ: increased permeability of microperoxidase MP-11 and horseradish peroxidase type VI	^Citation364 ^Citation9
Par-C5  = Pa-4 rat	-parotid gland acinar cells -replication defective SV40 immortalization	DMEM F-12 (1:1) + 2.5% FCS + 5 µg/ml insulin + 5 µg/ml transferrin + 25 ng/ml EGF + 1.1 µM hydrocortisone + 5 mM glutamate + 60 µg/ml kanamycin monosulfate^14	n.a. n.a. n.a.	-Transwell, collagen-coated clear PC filters, 1cm^2 growth area, 0.4 µm pore size, Costar, 10^6 cells/cm^2 Clearwell, Costar Corning	-TEM: secretory granules, TJ, intermediate junctions, desmosomes, microvilli -TEER^Δ: 2000 Ωcm^2 (d7) -TJ/AJ expression: IF (d n.i.): Occ^pos, ZO-1^pos, E-cad^pos WB (d n.i.): Occ, ZO-1, E-cad, Cldn1 -TEER^Δ: ~ 2000 Ωcm^2 (d5) -TJ/AJ expression: IF (d n.i.): β-cat^pos, Occ^pos	^Citation364 ^Citation343 ^Citation344

d … days in culture, n.a. … not analyzed, n.i. … not indicated, n.d. … not detected, f.c. … final concentration

# … listed as cross-contaminated cell lines (HeLa derivates by ATCC, ECACC

Δ … TEER readings from filters without cells were subtracted from values obtained with filters and seeded cells

1) establishment and characterization in different cell culture medium: MEM, 10% FCS, 2 mmol/l L-glutamine ^365,366)

2) different antibiotic/antimycotic substances used: 10 mg/ml gentamicin instead of 2.5 µg/ml amphotericin B³⁴⁷

3) for Transwell inserts, membrane diameters are given in mm

4) collagen I/III coating of filters had no effect on TEER

5) CSG WT monolayers used as control cell line, i.e. inulin flux set to 100%

6) for all experiments, cells were used at the second passage, 5 – 7 days after plating

7) Cldn3 main claudin expressed in SMIE cells

8) for FITC-dextran flux, medium was changed to HEPES-buffered saline (10 mM HEPES/NaOH pH 7.4, 1 mM sodium pyruvate, 10 mM glucose, 3 mM CaCl2, 145 mM NaCl)

9) for FITC-dextran flux, medium was changed to DMEM containing 10 mM HEPES buffer (pH 7.4)

10) WT SMG-C6 monolayers given as “relative paracellular flux” = 1

11) TEER and paracellular flux given for scrambled siRNA SMG-C6 control cells, i.e. relative paracellular flux of FITC-dextran set to 1

12) apparent permeability coefficients given as “% of initial value”, WT SMG-C6 set to 100%

13) peroxidase activity in cell supernatant given as “absorbance values at 405nm”

14) Li et al.³⁴³ used 600 µg/ml G418 instead of kanamycin monosulfate; Quissel et al.³⁶⁴ additionally used 8.4 ng/ml cholera toxin

abbreviations: αAMY, α amylase; αSMA, α smooth muscle actin; AICAR, 5-aminoimidazole-4-carboxamideribonucleoside (AMPK activator); AJ, adherens junction; AMPK, AMP-activated protein kinase; AQP, aquaporin; β-cat, β-catenin; BEBM, bronchial epithelial basal medium; CAP, capsaicin; Cch, carbachol; CK; cytokeratin (panel); Cldn, claudin; DMEM, Dulbecco’s Modified Eagle Medium; E-cad, E – cadherin; EGF, epidermal growth factor; fAd, full length adiponectin; FCS, fetal calf serum; gAd, globular adiponectin; hTERT, human telomerase reverse transcriptase; IF, immunofluorescence; JAM-A, junctional adhesion molecule A; MEM, minimum essential media; MG, Matrigel; MUC, mucin; Occ, occludin; PC, polycarbonate; PET, polyester; TEM, transmission electron microscopy; TEER, transepithelial electrical resistance; TJ, tight junction; VIM, vimentin; WB, Western Blot; ZO, zonula occludens;

Table 5. Salivary gland primary epithelial cells: This table summarizes primary cell culture techniques that have been used for the study of epithelial barrier properties of salivary glands.

Primary Cells	Cell Culture Medium	Max. Passage used	Characterization (ductal/acinar)	Model Set-Up (permeability assays)	Studied Barrier Properties	References
phmSG human minor salivary gland	KGM-high (0.8 nM Ca^2+) + BPEs + hEGF + insulin + hydrocortisone + gentamicin + epinephrine + transferrin^1	8–10 (in KGM-low, 0.05 nM Ca^2+)	-PCR (d3)^2: ZO-1^high, CLDN1^high, AQP5^rel.high, SLC12A2 ^rel.high, CST3^rel.high, ORAI1^rel.high, αAMY^modest, ANO1^modest KRT19^low, KLK1^low -IF (d3): SLC12A2^pos, CSTe^pos, AQP5^pos -agonist-induced αAMY secretion and activity	Transwell-COL, collagen-coated PET filter, 0.4 µm pore size, Corning, 20 × 10^4 cells/well	-TER^Δ3: ~ 90–180 Ωcm^2 (time-course, d3-d10) -TJ/AJ expression: IF (d3): ZO-1^pos PCR (d3):E-cad, ZO-1, Cldn1	^Citation367
huSMG human submandibular gland	serum-free Hepato-STIM + 1% glutamine + 500 U/ml penicillin + 500 µg/ml streptomycin + 12.5 µg/ml fungizone	n.i. 3	-IF (d n.i.): E-cad^pos, Vim^neg -PCR (d n.i.): αAMY, AQP5, NKCC1, ENaC, Cldn1 - αAMY secretion: strong secretion on Transwell vs plastic (5022–1682 U/l vs 742–214 U/l)	24 mm Transwell-Clear, PET filter, Corning Transwell-Clear, PET filters, FCS-coated, 0.4 µm pore size, 1.12 cm^2 growth area, Corning Costar	-TER^Δ: ~ 250–300 Ωcm^2 (d n.i.) -TJ/AJ expression: IF (d n.i.): ZO-1^pos, Cldn1^pos, E-cad^pos -TEM: tight junctions, microvilli, no secretory granules -TER^Δ: 622 ± 117 Ωcm^2 (d7-d10)^4 -TJ/AJ expression: PCR (d n.i.): Cldn1	^Citation368 ^Citation369 5
huSG human submandibular or parotid gland	serum-free Hepato-STIM + 1% glutamine + 500 U/ml penicillin + 500 µg/ml streptomycin + 12.5 µg/ml fungizone	3	-IF (d n.i.): non-coated: CK^pos(85%), VIM^mod(40%), αSMA^neg, αAMY^neg, AQP5^neg, MUC1^neg MG-coated: CK^pos, VIM^pos, αSMA^neg, αAMY^pos, AQP5^pos, MUC1^mod(50%) -enhanced αAMY secretion in MG-coated set-up	24 mm Transwell-Clear, non-coated or MG-coated (f.c. 2 mg/mL BD Biosciences) PET filters, Corning, 5 × 10^4 cells/cm^2	-TER^Δ: non-coated: 381, 409, 415 Ωcm^2 (d3, d5, d7); MG-coated: 409, 407, 415 Ωcm^2 (d3, d5, d7) -TJ/AJ expression: IF (d n.i.): non-coated± MG-coated: Cldn1^pos, Cldn2^pos, Cldn3^pos, Cldn4^pos, Occ^pos, JAM-A^pos, ZO-1^pos -TEM: non-coated: TJs, no Golgi apparatus, no rER, no secretory granules, no microvilli MG-coated: TJs, many Golgi saccules, rER, secretory granules	^Citation370
PTHSG^6 human submandibular gland	Hepato-STIM + 10% FCS + 1% glutamine + 500 U/ml penicillin + 500 µg/ml streptomycin + 12.5 µg/ml fungizone	4	IF (p0): non-coated or BME-coated: αAMY^pos and αAMY^neg clusters, non-coated: some Vim^pos clusters, BME-coated: Vim^neg	-	-TJ/AJ expression: PCR: describes differential expression of αAMY, Cldn1, Cldn3, KLK and Vim in 13 patients	^Citation371
		3	-PCR (d n.i.): αAMY, AQP5, NKCC1, ENaC, Cldn1 - αAMY secretion: strong secretion on Transwell vs plastic (3043–882 U/l vs 568–193 U/l)	Transwell-Clear, PET filters, FCS-coated, 0.4 µm pore size, 1.12 cm^2 growth area, Corning Costar	-TER^Δ: 595 ± 93 Ωcm^2 (d7-d10)^4 -TJ/AJ expression: PCR (d n.i.): Cldn1	^Citation369
RPG rhesus monkey parotid gland	serum-free Hepato-STIM + 1% glutamine + 500 U/ml penicillin + 500 µg/ml streptomycin + 12.5 µg/ml fungizone	4	IF (d n.i.): E-cad^pos, Na/K ATPase^pos	24 mm Transwell-Clear, PET filter, Corning	-TER^Δ: ~ 135 Ωcm^2 (d3) -TJ/AJ expression: IF (d n.i.): E-cad^pos, ZO-1^pos, Cldn1^pos -restricted paracellular fluid movement (2.0 µl/cm^2 vs. 6.5 µl/cm^2 in HSG)	^Citation372
ASGE BALB/c mouse submandibular gland	DMEM–F12 + 2 mM glutamine + 10 ng/ml EGF + 5 µg/ml insulin + 5 µg/ml transferrin + 100 U/ml penicillin + 100 U/ml streptomycin + 0.25 µg/ml amphotericin + 2 nM triiodothyronine T3 + 0.4 µg/ml hydrocortisone + 0.18 mM adenine + 100 nM cholera toxin	5	IF (d n.i.): CK^pos, VIM^neg (ASGEs on feeder layer of irradiated, VIM^pos NIH fibroblasts)	24 mm Transwell, PC filters, Corning Costar, 1 × 10^6 cells/well	-TER^Δ: ~ 150 Ωcm^2 (d5) -TEM: desmosomes, TJs, microvilli, secretory granules (ASGEs on feeder layer of irradiated NIH 3T3 fibroblasts)	^Citation373

d … days in culture, n.a. … not analyzed, n.i. … not indicated, n.d. … not detected, f.c. … final concentration

Δ … TEER readings from filters without cells were subtracted from values obtained with filters and seeded cells

1 Jang et al.³⁶⁷ tested 6 differently supplemented growth media (no concrete supplement concentrations given). In this table, growth conditions that achieve the maintenance of phmSG in an acinar-like phenotype are given.

2) Expression levels of respective genes (high, relatively high, modest, low; in comparison to expression levels in other growth media) were adopted from Jang et al..³⁶⁷

3) Cells grown in KGM-H (0.8 nM Ca²⁺) showed fastest and highest rates of TEER development; after 7 days, TEER showed no significant difference between cells grown in KGM-low and KGM-high.

4) In MEM medium, TEER values were below 200 Ωcm².

5) Preparation of tissue biopsies and cell isolation as previously described by Tran et al. (2005), with some modifications.

6) The isolation and culture method for PTHSGs described by Szlavik et al.³⁷¹ and Hegyesi et al.³⁶⁹ is based on the protocol of Tran et al.,³⁶⁸ with some modifications.

abbreviations: αAMY, alpha amylase; AQP, aquaporin; BPEs, bovine pituitary extracts; β-cat, β-catenin; CK; cytokeratin (panel); Cldn, claudin; d, day; DMEM, Dulbecco’s Modified Eagle Medium; E-cad, E – cadherin; EGF, epidermal growth factor; FCS, fetal calf serum; FH, fibrin hydrogel; GFR, growth factor reduced; hTERT, human telomerase reverse transcriptase; JAM-A, junctional adhesion molecule A; KGM, keratinocyte growth medium; MG, Matrigel; MUC, mucin; n.a., not analyzed; Occ, occludin; PC, polycarbonate; PET, polyester; P_app, apparent permeability coefficient; rER, rough endoplasmic reticulum; VIM, vimentin; ZO, zonula occludens;

Rupniak HT, et al. Characteristics of four new human cell lines derived from squamous cell carcinomas of the head and neck. J Natl Cancer Inst. 1985;75(4):621–35.

 PubMedGoogle Scholar

Klemetsrud T, et al. Polymer coated liposomes for use in the oral cavity - A study of the in vitro toxicity, effect on cell permeability and interaction with mucin. J Liposome Res. 2016;3:1–37.

 Google Scholar

Pistone S, et al. Formulation of polysaccharide-based nanoparticles for local administration into the oral cavity. Eur J Pharm Sci. 2016;96:381–9. doi:10.1016/j.ejps.2016.10.012.

 PubMed Web of Science ®Google Scholar

Zeng N, et al. Poloxamer bioadhesive hydrogel for buccal drug delivery: Cytotoxicity and trans-epithelial permeability evaluations using TR146 human buccal epithelial cell line. Int J Pharm. 2015;495(2):1028–37. doi:10.1016/j.ijpharm.2015.09.045.

 PubMed Web of Science ®Google Scholar

Moyes DL, et al. A biphasic innate immune MAPK response discriminates between the yeast and hyphal forms of Candida albicans in epithelial cells. Cell Host Microbe. 2010;8(3):225–35. doi:10.1016/j.chom.2010.08.002.

 PubMed Web of Science ®Google Scholar

Hiroshima Y, et al. Effect of Hangeshashinto on calprotectin expression in human oral epithelial cells. Odontology. 2016;104(2):152–62. doi:10.1007/s10266-015-0196-3.

 PubMed Web of Science ®Google Scholar

Lermann U, Morschhäuser J. Secreted aspartic proteases are not required for invasion of reconstituted human epithelia by Candida albicans. Microbiology. 2008;154(Pt 11):3281–95. doi:10.1099/mic.0.2008/022525-0.

 PubMedGoogle Scholar

Reiss M, Pitman SW, Sartorelli AC. Modulation of the terminal differentiation of human squamous carcinoma cells in vitro by all-trans-retinoic acid. J Natl Cancer Inst. 1985;74(5):1015–23.

 PubMedGoogle Scholar

Hansson A, et al. Expression of keratins in normal, immortalized and malignant oral epithelia in organotypic culture. Oral Oncol. 2001;37(5):419–30. doi:10.1016/S1368-8375(00)00089-0.

 PubMed Web of Science ®Google Scholar

Sarang Z, et al. Microarray assessment of fibronectin, collagen and integrin expression and the role of fibronectin-collagen coating in the growth of normal, SV40 T-antigen-immortalised and malignant human oral keratinocytes. Altern Lab Anim. 2003;31(6):575–85.

 PubMed Web of Science ®Google Scholar

Chun SG, et al. Targeted inhibition of histone deacetylases and hedgehog signaling suppress tumor growth and homologous recombination in aerodigestive cancers. Am J Cancer Res. 2015;5(4):1337–52.

 PubMed Web of Science ®Google Scholar

Moroyama T, et al. Establishment and characterization of a human tumor cell line (Nakata-1) derived from squamous carcinoma of the buccal mucosa. Proc Jpn Can Assoc. 1986;45: 242.not available.

 Google Scholar

Sato K, et al. RANKL synthesized by both stromal cells and cancer cells plays a crucial role in osteoclastic bone resorption induced by oral cancer. Am J Pathol. 2013;182(5):1890–9. doi:10.1016/j.ajpath.2013.01.038.

 PubMed Web of Science ®Google Scholar

Laheij AM, et al. The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro. J Oral Microbiol. 2015;7:27543. doi:10.3402/jom.v7.27543.

 PubMed Web of Science ®Google Scholar

Hayashi K, et al. Effect of 9-cis-retinoic acid on oral squamous cell carcinoma cell lines. Cancer Lett. 2000;151(2):199–208. doi:10.1016/S0304-3835(99)00422-X.

 PubMed Web of Science ®Google Scholar

Hayashi K, et al. Overexpression of retinoic acid receptor beta induces growth arrest and apoptosis in oral cancer cell lines. Jpn J Cancer Res. 2001;92(1):42–50. doi:10.1111/j.1349-7006.2001.tb01046.x.

 PubMedGoogle Scholar

Prime SS, et al. The behaviour of human oral squamous cell carcinoma in cell culture. J Pathol. 1990;160(3):259–69. doi:10.1002/path.1711600313.

 PubMed Web of Science ®Google Scholar

Yee M, et al. Porphyromonas gingivalis stimulates IL-6 and IL-8 secretion in GMSM-K, HSC-3 and H413 oral epithelial cells. Anaerob. 2014;28:62–7. doi:10.1016/j.anaerobe.2014.05.011.

 PubMed Web of Science ®Google Scholar

Narayanan SP, et al. Integrated genomic analyses identify KDM1A’s role in cell proliferation via modulating E2F signaling activity and associate with poor clinical outcome in oral cancer. Cancer Lett. 2015;367(2):162–72. doi:10.1016/j.canlet.2015.07.022.

 PubMed Web of Science ®Google Scholar

Ye P, et al. Differential expression of transforming growth factors-beta 1, -beta 2, -beta 3 and the type I, II, III receptors in the lining epithelia of inflamed gingiva. Pathology. 2003;35(5):384–92. doi:10.1080/00313020310001602585.

 PubMed Web of Science ®Google Scholar

Lee EJ, et al. Characterization of newly established oral cancer cell lines derived from six squamous cell carcinoma and two mucoepidermoid carcinoma cells. Exp Mol Med. 2005;37(5):379–90. doi:10.1038/emm.2005.48.

 PubMed Web of Science ®Google Scholar

Kim YK, Koo NY, Yun PY. Anticancer effects of CKD-602 (Camtobell®) via G2/M phase arrest in oral squamous cell carcinoma cell lines. Oncol Lett. 2015;9(1):136–42. doi:10.3892/ol.2014.2648.

 PubMed Web of Science ®Google Scholar

Moon, Y, et al. Effect of CD133 overexpression on the epithelial-to-mesenchymal transition in oral cancer cell lines. Clin Exp Metastasis. 2016;33(5):487–96. doi:10.1007/s10585-016-9793-y.

 PubMed Web of Science ®Google Scholar

Lin, SC, et al. Establishment of OC3 oral carcinoma cell line and identification of NF-kappa B activation responses to areca nut extract. J Oral Pathol Med. 2004;33(2);79–86. doi:10.1111/j.1600-0714.2004.00034.x.

 PubMed Web of Science ®Google Scholar

Chen, YH, et al. Apoptotic effect of cisplatin and cordycepin on OC3 human oral cancer cells. Chin J Integr Med. 2014;20(8):624–32. doi:10.1007/s11655-013-1453-3.

 PubMed Web of Science ®Google Scholar

Wu, SY, Wu, AT, Liu, SH. Microrna-17-5p regulated apoptosis-related protein expression and radiosensitivity in oral squamous cell carcinoma caused by betel nut chewing. Oncotarget. 2016;7(32):51482–51493. doi:10.18632/oncotarget.9856.

 Google Scholar

Hsiao, JR, Leu, SF, Huang, BM. Apoptotic mechanism of paclitaxel-induced cell death in human head and neck tumor cell lines. J Oral Pathol Med. 2009;38(2):188–97. doi:10.1111/j.1600-0714.2008.00732.x.

 PubMed Web of Science ®Google Scholar

Wang, SH, Chang, CW, Chou, HC. 5-Methoxytryptophan-dependent inhibition of oral squamous cell carcinoma metastasis. Electrophoresis. 2015;36(17), pp. 2027–34. doi:10.1002/elps.201500154.

 PubMed Web of Science ®Google Scholar

Kok, SH, et al. Establishment and characterization of a tumorigenic cell line from areca quid and tobacco smoke-associated buccal carcinoma. Oral Oncol. 2007;43(7):639–47. doi:10.1016/j.oraloncology.2006.07.007.

 PubMed Web of Science ®Google Scholar

Lin, CS, et al. Silencing JARID1B suppresses oncogenicity, stemness and increases radiation sensitivity in human oral carcinoma. 2015;368(1);36–45.

 Google Scholar

Lee, CH, et al. IL-1β promotes malignant transformation and tumor aggressiveness in oral cancer. J Cell Physiol. 2015;230(4):875–84. doi:10.1002/jcp.24816.

 Web of Science ®Google Scholar

Chang, CC, et al. HDAC2 promotes cell migration/invasion abilities through HIF-1α stabilization in human oral squamous cell carcinoma. J Oral Pathol Med. 2011;40(7):567–75. doi:10.1111/j.1600-0714.2011.01009.x.

 Web of Science ®Google Scholar

Gioanni, J, et al. Two new human tumor cell lines derived from squamous cell carcinomas of the tongue: establishment, characterization and response to cytotoxic treatment. Eur J Cancer Clin Oncol. 1988;24(9);1445–55. doi:10.1016/0277-5379(88)90335-5.

 PubMedGoogle Scholar

Zhang, Z, et al. Hyaluronan synthase 2 expressed by cancer-associated fibroblasts promotes oral cancer invasion. 2016;35(1):181. doi:10.1186/s13046-016-0458-0.

 Google Scholar

Li, YC, et al. SATB1 promotes tumor metastasis and invasiveness in oral squamous cell carcinoma. Oral Dis. 2016; 23(2):247–254. doi:10.1111/odi.12602.

 Google Scholar

Zhang, X, et al. Knockdown of Myosin 6 inhibits proliferation of oral squamous cell carcinoma cells. J Oral Pathol Med. 2016;45(10):740–5. doi:10.1111/jop.12448.

 PubMed Web of Science ®Google Scholar

Bozec, A, et al. Combination of phosphotidylinositol-3-kinase targeting with cetuximab and irradiation: A preclinical study on an orthotopic xenograft model of head and neck cancer. Head Neck. 2017;39(1):151–159.  doi:10.1002/hed.24560.

 Google Scholar

Stojanović, N, et al. Integrin αvβ3 expression in tongue squamous carcinoma cells Cal27 confers anticancer drug resistance through loss of pSrc(Y418). Biochim Biophys Acta. 2016;1863(8):1969–78. doi:10.1016/j.bbamcr.2016.04.019.

 PubMed Web of Science ®Google Scholar

Mo, H, et al. Expression, roles and therapy target values of CD24 in oral squamous cell carcinoma. Beijing Da Xue Xue Bao. 2016;48(1):16–22. Article in Chinese.

 Google Scholar

Ma, Z, Bi, Q and Wang, Y. Hydrogen sulfide accelerates cell cycle progression in oral squamous cell carcinoma cell lines. Oral Dis. 2015;21(2):156–62. doi:10.1111/odi.12223.

 PubMed Web of Science ®Google Scholar

Wu, HH, et al. Bevacizumab regulates cancer cell migration by activation of STAT3. Asian Pac J Cancer Prev. 2015;16(5):6501–6. doi:10.7314/APJCP.2015.16.15.6501.

 PubMedGoogle Scholar

Cheung, J, et al. A nonviral vector with transfection activity comparable with adenoviral transduction. Ther Deliv. 2016;7(11):739–49. doi:10.4155/tde-2016-0054.

 PubMed Web of Science ®Google Scholar

Samal, SK, et al. Ketorolac salt is a newly discovered DDX3 inhibitor to treat oral cancer. Sci Rep. 2015;5:9982. doi:10.1038/srep09982.

 PubMed Web of Science ®Google Scholar

Rheinwald, JG and Beckett, MA. Tumorigenic keratinocyte lines requiring anchorage and fibroblast support cultured from human squamous cell carcinomas. Cancer Res. 1981;41(5):1657–63.

 PubMed Web of Science ®Google Scholar

Otsuka, Y, et al. High expression of EPB41L5, an integral component of the Arf6-driven mesenchymal program, correlates with poor prognosis of squamous cell carcinoma of the tongue. Cell Commun Signal. 2016;14(1):28. doi:10.1186/s12964-016-0151-0.

 PubMedGoogle Scholar

Kadletz, L, et al. AZD5582, an IAP antagonist that leads to apoptosis in head and neck squamous cell carcinoma cell lines and is eligible for combination with irradiation. Acta Otolaryngol. 2016; 137(3):320–325. doi:10.1080/00016489.2016.1242776.

 Google Scholar

McGregor, F, et al. Molecular changes associated with oral dysplasia progression and acquisition of immortality: potential for its reversal by 5-azacytidine. Cancer Res. 2002;62(16):4757–66.

 PubMed Web of Science ®Google Scholar

Colley, HE, et al. Development of tissue-engineered models of oral dysplasia and early invasive oral squamous cell carcinoma. Br J Cancer. 2011;105(10):1582–92. doi:10.1038/bjc.2011.403.

 PubMed Web of Science ®Google Scholar

Mian, SA, et al. Raman spectroscopy can discriminate between normal, dysplastic and cancerous oral mucosa: a tissue-engineering approach. J Tissue Eng Regen Med. 2016; 11(11):3253–3262. doi:10.1002/term.2234.

 Google Scholar

Chang, SE, et al. DOK, a cell line established from human dysplastic oral mucosa, shows a partially transformed non-malignant phenotype. Int J Cancer. 1992;52(6):896–902. doi:10.1002/ijc.2910520612.

 PubMed Web of Science ®Google Scholar

Gaballah, K, et al. Tissue engineering of oral dysplasia. J Pathol. 2008;215(3):280–9. doi:10.1002/path.2360.

 PubMed Web of Science ®Google Scholar

Kulasekara, KK, et al. Cancer progression is associated with increased expression of basement membrane proteins in three-dimensional in vitro models of human oral cancer. Arch Oral Biol. 2009;54(10):924–31. doi:10.1016/j.archoralbio.2009.07.004.

 PubMed Web of Science ®Google Scholar

Chan LP, et al. IL-8 promotes HNSCC progression on CXCR1/2-meidated NOD1/RIP2 signaling pathway. Oncotarget. 2016; 7(38):61820–61831. doi:10.18632/oncotarget.11445.

 Google Scholar

Dong Y, et al. Establishment of a new OSCC cell line derived from OLK and identification of malignant transformation-related proteins by differential proteomics approach. Sci Rep. 2015;5:12668. doi:10.1038/srep12668.

 PubMed Web of Science ®Google Scholar

Rikimaru, K, et al. Growth of the malignant and nonmalignant human squamous cells in a protein-free defined medium. In Vitro Cell Dev Biol. 1990;26(9):849–56. doi:10.1007/BF02624609.

 PubMed Web of Science ®Google Scholar

Harada, K, et al. Metformin in combination with 5-fluorouracil suppresses tumor growth by inhibiting the Warburg effect in human oral squamous cell carcinoma. Int J Oncol. 2016;49(1):276–84. doi:10.3892/ijo.2016.3523.

 PubMed Web of Science ®Google Scholar

Ketkaew, Y, et al. Apigenin inhibited hypoxia induced stem cell marker expression in a head and neck squamous cell carcinoma cell line. Arch Oral Biol. 74:69–74. doi:10.1016/j.archoralbio.2016.11.010.

 Google Scholar

Momose, F, et al. Variant sublines with different metastatic potentials selected in nude mice from human oral squamous cell carcinomas. J Oral Pathol Med. 1989;18(7):391–5. doi:10.1111/j.1600-0714.1989.tb01570.x.

 PubMed Web of Science ®Google Scholar

Yuan, Z, et al. Overexpression of HOXB7 protein reduces sensitivity of oral cancer cells to chemo-radiotherapy. Cancer Gene Ther. 2016;23:12:419–24. doi:10.1038/cgt.2016.55.

 PubMed Web of Science ®Google Scholar

Kaneko, T, et al. Hypoxia-induced epithelial-mesenchymal transition is regulated by phosphorylation of GSK3-β via PI3 K/Akt signaling in oral squamous cell carcinoma. Oral Surg Oral Med Oral Pathol Oral Radiol. 2016;122(6):719–s730. doi:10.1016/j.oooo.2016.06.008.

 PubMed Web of Science ®Google Scholar

Yokoi, T, Homma, H, Odajima, T. Establishment and characterization of OSC-19 cell line in serum and protein free culture. Tumor Res. 1988;24:1–17.

 Google Scholar

Shibata, A, et al. Synthetic terrein inhibits progression of head and neck cancer by suppressing angiogenin production. Anticancer Res. 2016;36(5):2161–8.

 PubMed Web of Science ®Google Scholar

Kitahara, H, et al. Eribulin sensitizes oral squamous cell carcinoma cells to cetuximab via induction of mesenchymal-to-epithelial transition. Oncol Rep. 2016;36(6);3139–44. doi:10.3892/or.2016.5189.

 PubMed Web of Science ®Google Scholar

Yokoi, T, et al. Some properties of a newly established human cell line derived from an oral squamous carcinoma. Tumor Res. 1990;25(93):91–93.

 Google Scholar

Kawasaki, G, et al. Overexpression of metastasis-associated MTA1 in oral squamous cell carcinomas: correlation with metastasis and invasion. Int J Oral Maxillofac Surg. 2008;37(11):1039–46. doi:10.1016/j.ijom.2008.05.020.

 PubMed Web of Science ®Google Scholar

Todoroki, K, et al. CD44v3+/CD24- cells possess cancer stem cell-like properties in human oral squamous cell carcinoma. Int J Oncol. 2016;48(1):99–109. doi:10.3892/ijo.2015.3261.

 PubMed Web of Science ®Google Scholar

Urade, M, et al. Establishment of human squamous carcinoma cell lines highly and minimally sensitive to bleomycin and analysis of factors involved in the sensitivity. Cancer. 1992;69(10):2589–97. doi:10.1002/1097-0142(19920515)69:10%3c2589:: AID-CNCR2820691032%3e3.0.CO;2-Y.

 PubMed Web of Science ®Google Scholar

Yazama, H, et al. Dietary glucosylceramides suppress tumor growth in a mouse xenograft model of head and neck squamous cell carcinoma by the inhibition of angiogenesis through an increase in ceramide. Int J Clin Oncol. 2015;20(3):438–46. doi:10.1007/s10147-014-0734-y.

 PubMed Web of Science ®Google Scholar

Kitano, H, et al. Long-term gene therapy with Del1 fragment using nonviral vectors in mice with explanted tumors. Onco Targets Ther. 2016;9:503–16.

 PubMed Web of Science ®Google Scholar

Berndt, A, et al. Oral squamous cell carcinoma invasion is associated with a laminin-5 matrix re-organization but independent of basement membrane and hemidesmosome formation. clues from an in vitro invasion model. Invasion Metastasis. 1997;17(5):251–8.

 PubMedGoogle Scholar

Dalley, AJ, et al. Organotypic culture of normal, dysplastic and squamous cell carcinoma-derived oral cell lines reveals loss of spatial regulation of CD44 and p75 NTR in malignancy. J Oral Pathol Med. 2013;42(1):37–46. doi:10.1111/j.1600-0714.2012.01170.x.

 PubMed Web of Science ®Google Scholar

Palmerini, CA, et al. Synthesis of new indole-based bisphosphonates and evaluation of their chelating ability in PE/CA-PJ15 cells. Eur J Med Chem. 2015;102:403–12. doi:10.1016/j.ejmech.2015.08.019.

 PubMed Web of Science ®Google Scholar

Büttner, R, et al. Myofibroblasts have an impact on expression, dimerization and signaling of different ErbB receptors in OSCC cells. J Recept Signal Transduct Res. 2016;37(1):25–37. doi:10.3109/10799893.2016.1155066.

 Google Scholar

Jo, JR, Park, YK, Jang, BC. Short-term treatment with glucosamine hydrochloride specifically downregulates hypoxia-inducible factor-1α at the protein level in YD-8 human tongue cancer cells. Int J Oncol. 2014;44(5):1699–706. doi:10.3892/ijo.2014.2336.

 PubMed Web of Science ®Google Scholar

So, KY, Oh, SH. Heme oxygenase-1-mediated apoptosis under cadmium-induced oxidative stress is regulated by autophagy, which is sensitized by tumor suppressor p53. Biochem Biophys Res Commun. 2016;479(1):80–5. doi:10.1016/j.bbrc.2016.09.037.

 PubMed Web of Science ®Google Scholar

Jung, CW, et al. Anti-cancer properties of glucosamine-hydrochloride in YD-8 human oral cancer cells: Induction of the caspase-dependent apoptosis and down-regulation of HIF-1α. Toxicol In Vitro. 2012;26(1):42–50. doi:10.1016/j.tiv.2011.10.005.

 PubMed Web of Science ®Google Scholar

Ahn, SG, et al. The anticancer mechanism of 2’-hydroxycinnamaldehyde in human head and neck cancer cells. Int J Oncol. 2015;47(5):1793–800. doi:10.3892/ijo.2015.3152.

 PubMed Web of Science ®Google Scholar

Anh, TD, et al. The histone deacetylase inhibitor, Trichostatin A, induces G2/M phase arrest and apoptosis in YD-10B oral squamous carcinoma cells. Oncol Rep. 2012:27(2):455–60.

 PubMed Web of Science ®Google Scholar

Yu, HJ, et al. Inhibition of myeloid cell leukemia-1: Association with sorafenib-induced apoptosis in human mucoepidermoid carcinoma cells and tumor xenograft. Head Neck. 2015;37(9):1326–35. doi:10.1002/hed.23749.

 PubMed Web of Science ®Google Scholar

Lim, W, et al. Association between cancer stem cell-like properties and epithelial-to-mesenchymal transition in primary and secondary cancer cells. 2016;49(3):991–1000.

 Google Scholar

Rheinwald, JG, Beckett, MA. Defective terminal differentiation in culture as a consistent and selectable character of malignant human keratinocytes. Cell. 1980;22 (2.2):629–32. doi:10.1016/0092-8674(80)90373-6.

 Google Scholar

Sun, L, et al. MicroRNA-137 suppresses tongue squamous carcinoma cell proliferation, migration and invasion. Cell Prolif. 2016;49(5):628–35. doi:10.1111/cpr.12287.

 PubMed Web of Science ®Google Scholar

Su, CC, et al. Cantharidin induced oral squamous cell carcinoma cell apoptosis via the JNK-Regulated mitochondria and endoplasmic reticulum stress-related rignaling pathways. PLoS One. 2016;11(12):e0168095. doi:10.1371/journal.pone.0168095.

 PubMed Web of Science ®Google Scholar

Dai, D, et al. Inhibition of mTOR/eIF4E by anti-viral drug ribavirin effectively enhances the effects of paclitaxel in oral tongue squamous cell carcinoma. Biochem Biophys Res Commun. 2016;482(4):1259–1264. doi:10.1016/j.bbrc.2016.12.025.

 Google Scholar

Jin, L, et al. Icaritin induces mitochondrial apoptosis by up-regulating miR-124 in human oral squamous cell carcinoma cells. Biomed Pharmacother. 2016;85:287–295. doi:10.1016/j.biopha.2016.11.023.

 Google Scholar

Korvala, J, et al. MicroRNA and protein profiles in invasive versus non-invasive oral tongue squamous cell carcinoma cells in vitro. Exp Cell Res. 2016;350(1):9–18. doi:10.1016/j.yexcr.2016.10.015.

 Google Scholar

Zhang, H, et al. HMGA2 is associated with the aggressiveness of tongue squamous cell carcinoma cells. Oral Dis. 2016;23(2):255–264. doi:10.1111/odi.12608.

 Google Scholar

Gustafson, DL, et al. Dose scheduling of the dual VEGFR and EGFR tyrosine kinase inhibitor vandetanib (ZD6474, Zactima) in combination with radiotherapy in EGFR-positive and EGFR-null human head and neck tumor xenografts. Cancer Chemother Pharmacol. 2008;61(2):179–88. doi:10.1007/s00280-007-0460-5.

 PubMed Web of Science ®Google Scholar

Bais, MV, Kukuruzinska, M, Trackman, PC. Orthotopic non-metastatic and metastatic oral cancer mouse models. Oral Oncol. 2015;51(5):476–82. doi:10.1016/j.oraloncology.2015.01.012.

 PubMed Web of Science ®Google Scholar

Harada, K, Ferdous, T, Ueyama, Y. Gimeracil exerts radiosensitizing effects on oral squamous cell carcinoma cells in vitro and in vivo. Anticancer Res. 2016;36(11):5923–30. doi:10.21873/anticanres.11179.

 PubMed Web of Science ®Google Scholar

Ohnishi, Y, et al. Lapatinib-resistant cancer cells possessing epithelial cancer stem cell properties develop sensitivity during sphere formation by activation of the ErbB/AKT/cyclin D2 pathway. Oncol Rep. 2016;36(5):3058–64. doi:10.3892/or.2016.5073.

 PubMed Web of Science ®Google Scholar

Horikoshi, M, et al. A new human cell line derived from human carcinoma of the gingiva. I. Its establishment and morphological studies. Nihon Koku Geka Gakkai Zasshi. 1974;20(2):100–6. Article in Japanese.

 Google Scholar

Tsubaki, M, et al. The sensitivity of head and neck carcinoma cells to statins is related to the expression of their Ras expression status, and statin-induced apoptosis is mediated via suppression of the Ras/ERK and Ras/mTOR pathways. Clin Exp Pharmacol Physiol. 2016;44(2):222–234. doi:10.1111/1440-1681.12690.

 Google Scholar

Kudo, Y, et al. Establishment of an oral squamous cell carcinoma cell line with high invasive and p27 degradation activities from a lymph node metastasis. Oral Oncol. 2003;39(5):515–20. doi:10.1016/S1368-8375(03)00015-0.

 PubMed Web of Science ®Google Scholar

Kudo, Y, et al. Establishment and characterization of a spindle cell squamous carcinoma cell line. 2006;35(8):479–83.

 Google Scholar

Inagaki, T, et al. treatment, Establishment of human oral-cancer cell lines (KOSC-2 and -3) carrying p53 and c-myc abnormalities by geneticin. Int J Cancer. 1994;56(2):301–8. doi:10.1002/ijc.2910560226.

 PubMed Web of Science ®Google Scholar

Maemoto, S, et al. Mutational analysis of HRAS and KRAS genes in oral carcinoma cell lines. Odontology. 2012;100(2):149–55. doi:10.1007/s10266-011-0032-3.

 PubMed Web of Science ®Google Scholar

Uozumi, M, et al. Induction of S100A4 gene expression inhibits in vitro invasiveness of human squamous cell carcinoma, KOSC-3 cells. Cancer Lett. 2000;149(1–2):135–41. doi:10.1016/S0304-3835(99)00352-3.

 PubMed Web of Science ®Google Scholar

Chiu, KC, et al. Polarization of tumor-associated macrophages and Gas6/Axl signaling in oral squamous cell carcinoma. Oral Oncol. 2015;51(7):683–9. doi:10.1016/j.oraloncology.2015.04.004.

 PubMed Web of Science ®Google Scholar

Yu, XD, et al. Resveratrol inhibits oral squamous cell carcinoma through induction of apoptosis and G2/M phase cell cycle arrest. Tumour Biol. 2016;37(3):2871–7. doi:10.1007/s13277-015-3793-4.

 PubMed Web of Science ®Google Scholar

Yang, CY, Meng, CL. Regulation of PG synthase by EGF and PDGF in human oral, breast, stomach, and fibrosarcoma cancer cell lines. J Dent Res. 1994;73(8):1407–15. doi:10.1177/00220345940730080301.

 PubMed Web of Science ®Google Scholar

Chang, KP, et al. Overexpression of caldesmon is associated with lymph node metastasis and poorer prognosis in patients with oral cavity squamous cell carcinoma. Cancer. 2013;119(22):4003–11. doi:10.1002/cncr.28300.

 PubMed Web of Science ®Google Scholar

Shirako, Y, et al. Heterogeneous tumor stromal microenvironments of oral squamous cell carcinoma cells in tongue and nodal metastatic lesions in a xenograft mouse model. J Oral Pathol Med. 2015;44(9):656–68. doi:10.1111/jop.12318.

 PubMed Web of Science ®Google Scholar

Oyama, G, et al. Single nucleotide polymorphisms of mucosa-associated lymphoid tissue 1 in oral carcinoma cells and gingival fibroblasts. Odontology. 2013;101(2):150–5. doi:10.1007/s10266-012-0079-9.

 PubMed Web of Science ®Google Scholar

Dickson, MA, et al. Human keratinocytes that express hTERT and also bypass a p16(INK4a)-enforced mechanism that limits life span become immortal yet retain normal growth and differentiation characteristics. Mol Cell Biol. 2000;20(4):1436–47. doi:10.1128/MCB.20.4.1436-1447.2000.

 PubMed Web of Science ®Google Scholar

Dalley, AJ, et al. Expression of ABCG2 and Bmi-1 in oral potentially malignant lesions and oral squamous cell carcinoma. Cancer Med. 2014;3(2):273–83. doi:10.1002/cam4.182.

 PubMed Web of Science ®Google Scholar

Miyauchi S, et al. Establishment of human tumor cell line (Ueda-1) derived from squamous cell carcinoma of the floor of the mouth. Jpn J Oral. Maxillofac. Surg. 1985;31:1347–51. in Japanese, not available. doi:10.5794/jjoms.31.1347.

 Google Scholar

Wang Y, et al. Evaluation of HO-1-u-1 cell line as an in vitro model for sublingual drug delivery involving passive diffusion–Initial validation studies. Int J Pharm. 2007; 334 (1–2): 27–34. doi:10.1016/j.ijpharm.2006.10.012.

 PubMedGoogle Scholar

Wang Y, Zuo Z, Chow MS. HO-1-u-1 model for screening sublingual drug delivery–influence of pH, osmolarity and permeation enhancer. Int J Pharm. 2009; 370 (1–2): 68–74. doi:10.1016/j.ijpharm.2008.11.010.

 PubMedGoogle Scholar

Saito, Y, et al. ALY as a potential contributor to metastasis in human oral squamous cell carcinoma. J Cancer Res Clin Oncol. 2013;139(4):585–94. doi:10.1007/s00432-012-1361-5.

 PubMed Web of Science ®Google Scholar

Roblegg, E, et al. Evaluation of a physiological in vitro system to study the transport of nanoparticles through the buccal mucosa. Nanotoxicology. 2012;6(4):399–413. doi:10.3109/17435390.2011.580863.

 PubMed Web of Science ®Google Scholar

Best, M, et al. Characterisation and cytotoxic screening of metal oxide nanoparticles putative of interest to oral healthcare formulations in non-keratinised human oral mucosa cells in vitro. Toxicol In Vitro. 2015;30(1.B):402–11. doi:10.1016/j.tiv.2015.09.022.

 Google Scholar

Gokulan, R, Halagowder, D. Expression pattern of Notch intracellular domain (NICD) and Hes-1 in preneoplastic and neoplastic human oral squamous epithelium: their correlation with c-Myc, clinicopathological factors and prognosis in Oral cancer. Med Oncol. 2014;31:8. doi:10.1007/s12032-014-0126-1.

 Web of Science ®Google Scholar

Ravindran, G, et al. Association of differential β-catenin expression with Oct- 4and Nanog in oral squamous cell carcinoma and their correlation with clinicopathological factors and prognosis. Head Neck. 2015;37(7):982–93. doi:10.1002/hed.23699.

 PubMed Web of Science ®Google Scholar

O’Neill, ID. Continued misrepresentation of KB cells as being of oral cancer phenotype requires action. Oral Oncol. 2009;45(10):e117–8. doi:10.1016/j.oraloncology.2009.02.005.

 PubMed Web of Science ®Google Scholar

Eagle, H. Propagation in a fluid medium of a human epidermoid carcinoma, strain KB. Proc Soc Exp Biol Med. 1955;89(3):362–4. doi:10.3181/00379727-89-21811.

 PubMed Web of Science ®Google Scholar

Karthikeyan, S, et al. Glaucarubinone sensitizes KB cells to paclitaxel by inhibiting ABC transporters via ROS-dependent and p53-mediated activation of apoptotic signaling pathways. Oncotarget. 2016;7(27):42353–73. doi:10.18632/oncotarget.9865.

 PubMedGoogle Scholar

Li, LK, et al. Goniothalamin induces cell cycle arrest and apoptosis in H400 human oral squamous cell carcinoma: A caspase-dependent mitochondrial-mediated pathway with downregulation of NF-κβ. Arch Oral Biol. 2016;64:28–38. doi:10.1016/j.archoralbio.2015.12.002.

 PubMed Web of Science ®Google Scholar

Shaghayegh, G, et al. Cell cycle arrest and mechanism of apoptosis induction in H400 oral cancer cells in response to Damnacanthal and Nordamnacanthal isolated from Morinda citrifolia. Cytotechnology. 2016;68(5):1999–2013. doi:10.1007/s10616-016-0014-y.

 PubMed Web of Science ®Google Scholar

Lourenço, SV, et al. Establishment and characterization of an oral mucosal melanoma cell line (MEMO) derived from a longstanding primary oral melanoma. Am J Dermatopathol. 2013;35(2):248–51. doi:10.1097/DAD.0b013e31826a9905.

 PubMed Web of Science ®Google Scholar

Sacks, PG, et al. Establishment and characterization of two new squamous cell carcinoma cell lines derived from tumors of the head and neck. Cancer Res. 1988;48(10):2858–66.

 PubMed Web of Science ®Google Scholar

Xi, S, et al. Decreased STAT1 expression by promoter methylation in squamous cell carcinogenesis. J Natl Cancer Inst. 2006;98(3):181–9. doi:10.1093/jnci/djj020.

 PubMedGoogle Scholar

Li, C, et al. Dasatinib blocks cetuximab- and radiation-induced nuclear translocation of the epidermal growth factor receptor in head and neck squamous cell carcinoma. Radiother Oncol. 2010;97(2):330–7. doi:10.1016/j.radonc.2010.06.010.

 PubMed Web of Science ®Google Scholar

Rangan, SR. A new human cell line (FaDu) from a hypopharyngeal carcinoma. Cancer. 1972;29(1):117–21. doi:10.1002/1097-0142(197201)29:1%3c117:: AID-CNCR2820290119%3e3.0.CO;2-R.

 PubMed Web of Science ®Google Scholar

Schneider, S, et al. Effects of neratinib and combination with irradiation and chemotherapy in head and neck cancer cells. Oral Dis. 2016;22(8):797–804. doi:10.1111/odi.12552.

 PubMed Web of Science ®Google Scholar

Chiu YW, et al. Tyrosine 397 phosphorylation is critical for FAK-promoted Rac1 activation and invasive properties in oral squamous cell carcinoma cells. Lab Invest. 2016;96(3):296–306. doi:10.1038/labinvest.2015.151.

 PubMed Web of Science ®Google Scholar

Kawamata H, et al. Possible contribution of active MMP2 to lymph-node metastasis and secreted cathepsin L to bone invasion of newly established human oral-squamous-cancer cell lines. Int J Cancer. 1997;70(1):120–7. doi:10.1002/(SICI)1097-0215(19970106)70:1%3c120:: AID-IJC18%3e3.0.CO;2-P.

 PubMed Web of Science ®Google Scholar

Tada T, et al. Oral squamous cell carcinoma cells modulate osteoclast function by RANKL-dependent and -independent mechanisms. Cancer Lett. 2009;274(1):126–31. doi:10.1016/j.canlet.2008.09.015.

 PubMed Web of Science ®Google Scholar

Kraus D, et al. Ghrelin promotes oral tumor cell proliferation by modifying GLUT1 expression. Cell Mol Life Sci. 2016;73(6):1287–99. doi:10.1007/s00018-015-2048-2.

 PubMed Web of Science ®Google Scholar

Harada K, et al. Induction of artificial cancer stem cells from tongue cancer cells by defined reprogramming factors. BMC Cancer. 2016;16, 548. doi:10.1186/s12885-016-2416-9.

 PubMed Web of Science ®Google Scholar

Ishisaki A, et al. Identification and characterization of autocrine-motility-factor-like activity in oral squamous-cell-carcinoma cells. Int J Cancer. 1994;59(6):783–8. doi:10.1002/ijc.2910590613.

 PubMed Web of Science ®Google Scholar

Nguyen PT, et al. The FGFR1 inhibitor PD173074 induces mesenchymal-epithelial transition through the transcription factor AP-1. Br J Cancer. 2013;109(8):2248–58. doi:10.1038/bjc.2013.550.

 PubMed Web of Science ®Google Scholar

Sakha S, et al. Exosomal microRNA miR-1246 induces cell motility and invasion through the regulation of DENND2D in oral squamous cell carcinoma. Sci Rep. 2016;6:38750. doi:10.1038/srep38750.

 PubMed Web of Science ®Google Scholar

Heo DS, et al. Biology, cytogenetics, and sensitivity to immunological effector cells of new head and neck squamous cell carcinoma lines. Cancer Res. 1989;49(18):5167–75.

 PubMed Web of Science ®Google Scholar

Minhas KM, et al. Spindle assembly checkpoint defects and chromosomal instability in head and neck squamous cell carcinoma. Int J Cancer. 2003;107(1):46–52. doi:10.1002/ijc.11341.

 PubMed Web of Science ®Google Scholar

Brenner JC, et al. Genotyping of 73 UM-SCC head and neck squamous cell carcinoma cell lines. Head Neck. 2010;32(4):417–26.

 PubMed Web of Science ®Google Scholar

Ganz T. Epithelia: not just physical barriers. Proc Natl Acad Sci U S A. 2002;99(6):3357–8. doi:10.1073/pnas.072073199.

 PubMed Web of Science ®Google Scholar

Kulkarni PS, et al. Characterization of human buccal epithelial cells transfected with the simian virus 40 T-antigen gene. Carcinogenesis. 1995;16(10):2515–21. doi:10.1093/carcin/16.10.2515.

 PubMed Web of Science ®Google Scholar

Hansson A, et al. Analysis of proliferation, apoptosis and keratin expression in cultured normal and immortalized human buccal keratinocytes. Eur J Oral Sci. 2003;111(1):34–41. doi:10.1034/j.1600-0722.2003.00010.x.

 PubMed Web of Science ®Google Scholar

Ceder R, et al. The application of normal, SV40 T-antigen-immortalised and tumour-derived oral keratinocytes, under serum-free conditions, to the study of the probability of cancer progression as a result of environmental exposure to chemicals. Altern Lab Anim. 2007;35(6):621–39.

 PubMed Web of Science ®Google Scholar

Ceder R, et al. Differentiation-promoting culture of competent and noncompetent keratinocytes identifies biomarkers for head and neck cancer. Am J Pathol. 2012;180(2):457–72. doi:10.1016/j.ajpath.2011.10.016.

 PubMed Web of Science ®Google Scholar

Chakravarti N, et al. Differential inhibition of protein translation machinery by curcumin in normal, immortalized, and malignant oral epithelial cells. Cancer Prev Res (Phila). 2010;3(3):331–8. doi:10.1158/1940-6207.CAPR-09-0076.

 PubMed Web of Science ®Google Scholar

Parikh N, et al. Isolation and characterization of an immortalized oral keratinocyte cell line of mouse origin. Arch Oral Biol. 2008;53(11):1091–100. doi:10.1016/j.archoralbio.2008.07.002.

 PubMed Web of Science ®Google Scholar

Conti HR, et al. IL-17 Receptor Signaling in Oral Epithelial Cells Is Critical for Protection against Oropharyngeal Candidiasis. Cell Host & Microbe. 2016;20(5):606–17. doi:10.1016/j.chom.2016.10.001.

 PubMed Web of Science ®Google Scholar

Takahashi C, et al. Newly established cell lines from mouse oral epithelium regenerate teeth when combined with dental mesenchyme. In Vitro Cell Dev Biol Anim. 2010;46(5):457–68. doi:10.1007/s11626-009-9265-7.

 PubMed Web of Science ®Google Scholar

Mäkelä M, Salo T, Larjava H. MMP-9 from TNF alpha-stimulated keratinocytes binds to cell membranes and type I collagen: a cause for extended matrix degradation in inflammation? Biochem Biophys Res Commun. 1998;253(2):325–35. doi:10.1006/bbrc.1998.9641.

 Google Scholar

Hildebrand HC, et al. Characterization of organotypic keratinocyte cultures on de-epithelialized bovine tongue mucosa. Histol Histopathol. 2002;17(1):151–63.

 PubMed Web of Science ®Google Scholar

Turunen A, et al. The combined effects of irradiation and herpes simplex virus type 1 infection on an immortal gingival cell line. Virol J. 2014;11, 125. doi:10.1186/1743-422X-11-125.

 PubMed Web of Science ®Google Scholar

Turunen A, Syrjänen S. Extracellular calcium regulates keratinocyte proliferation and HPV 16 E6 RNA expression in vitro. APMIS. 2014;122(9):781–9. doi:10.1111/apm.12227.

 PubMed Web of Science ®Google Scholar

Gursoy UK, et al. Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model. Cytotechnology. 2016;68(6):2345–54. doi:10.1007/s10616-016-0029-4.

 PubMed Web of Science ®Google Scholar

Oda D, et al. HPV immortalization of human oral epithelial cells: a model for carcinogenesis. Exp Cell Res. 1996;226(1):164–9. doi:10.1006/excr.1996.0215.

 PubMed Web of Science ®Google Scholar

Parikka M, et al. Alterations of collagen XVII expression during transformation of oral epithelium to dysplasia and carcinoma. J Histochem Cytochem. 2003;51(7):921–9. doi:10.1177/002215540305100707.

 PubMed Web of Science ®Google Scholar

Lee HJ, et al. Effects of nicotine on proliferation, cell cycle, and differentiation in immortalized and malignant oral keratinocytes. J Oral Pathol Med. 2005;34(7):436–43. doi:10.1111/j.1600-0714.2005.00342.x.

 PubMed Web of Science ®Google Scholar

Roesch-Ely M, et al. Organotypic co-cultures allow for immortalized human gingival keratinocytes to reconstitute a gingival epithelial phenotype in vitro. Differentiation. 2006;74(9–10):622–37. doi:10.1111/j.1432-0436.2006.00099.x.

 PubMed Web of Science ®Google Scholar

Buskermolen JK, et al. Development of a full-thickness human gingiva equivalent constructed from immortalized keratinocytes and fibroblasts. Tissue Eng Part C Methods. 2016;22(8):781–91. doi:10.1089/ten.tec.2016.0066.

 PubMed Web of Science ®Google Scholar

Kibe T, et al. Immortalization and characterization of normal oral epithelial cells without using HPV and SV40 genes. Oral Science International. 2011;8(1):20–28. doi:10.1016/S1348-8643(11)00009-7.

 Google Scholar

Hatakeyama S, et al. Establishment of gingival epithelial cell lines from transgenic mice harboring temperature sensitive simian virus 40 large T-antigen gene. J Oral Pathol Med. 2001;30(5):296–304. doi:10.1034/j.1600-0714.2001.300507.x.

 PubMed Web of Science ®Google Scholar

Hatakeyama S, et al. Retinoic acid disintegrated desmosomes and hemidesmosomes in stratified oral keratinocytes. J Oral Pathol Med. 2004;33(10):622–8. doi:10.1111/j.1600-0714.2004.00245.x.

 PubMed Web of Science ®Google Scholar

Hatakeyama S, Ishida K, Takeda Y. Changes in cell characteristics due to retinoic acid; specifically, a decrease in the expression of claudin-1 and increase in claudin-4 within tight junctions in stratified oral keratinocytes. J Periodontal Res. 2010;45(2):207–15. doi:10.1111/j.1600-0765.2009.01219.x.

 PubMed Web of Science ®Google Scholar

Hatakeyama S, et al. Expression of connexins and the effect of retinoic acid in oral keratinocytes. J Oral Sci. 2011;53(3):327–32. doi:10.2334/josnusd.53.327.

 PubMedGoogle Scholar

Miyata K, Takebe J. Anodized-hydrothermally treated titanium with a nanotopographic surface structure regulates integrin-α6β4 and laminin-5 gene expression in adherent murine gingival epithelial cells. J Prosthodont Res. 2013;57(2):99–108. doi:10.1016/j.jpor.2012.12.002.

 PubMed Web of Science ®Google Scholar

Nakayama Y, et al. Amelotin gene expression is temporarily being upregulated at the initiation of apoptosis induced by TGFβ1 in mouse gingival epithelial cells. Apoptosis. 2016;21(10):1057–70. doi:10.1007/s10495-016-1279-5.

 PubMed Web of Science ®Google Scholar

Moffatt-Jauregui, CE, et al. Establishment and characterization of a telomerase immortalized human gingival epithelial cell line. J Periodontal Res. 2013;48(6):713–21.

 PubMed Web of Science ®Google Scholar

Jauregui, CE, et al. Suppression of T-Cell Chemokines by Porphyromonas gingivalis. Infect Immun. 2013;81(7):2288–95. doi:10.1128/IAI.00264-13.

 PubMed Web of Science ®Google Scholar

Takeuchi, H, et al. The serine phosphatase SerB of Porphyromonas gingivalis suppresses IL-8 production by dephosphorylation of NF-κB RelA/p65. PLoS Pathog. 2013;9(4):e1003326. doi:10.1371/journal.ppat.1003326.

 PubMed Web of Science ®Google Scholar

Wang, H, et al. Porphyromonas gingivalis-induced reactive oxygen species activate JAK2 and regulate production of inflammatory cytokines through c-Jun. Infect Immun. 2014;82(10):4118–26. doi:10.1128/IAI.02000-14.

 PubMed Web of Science ®Google Scholar

Chen, SC, et al. MicroRNAs regulate cytokine responses in gingival epithelial cells. Infect Immun. 2016;84(12):3282–9. doi:10.1128/IAI.00263-16.

 PubMed Web of Science ®Google Scholar

Bao, K, Akguel, B, Bostanci, N. Establishment and characterization of immortalized gingival epithelial and fibroblastic cell lines for the development of organotypic cultures. Cells Tissues Organs. 2014;199(4):228–37. doi:10.1159/000363694.

 PubMed Web of Science ®Google Scholar

Kubo, C, et al. Immortalization of normal human gingival keratinocytes and cytological and cytogenetic characterization of the cells. Odontology. 2009;97(1):18–31. doi:10.1007/s10266-008-0089-9.

 PubMed Web of Science ®Google Scholar

Gröger S, Michel J, Meyle J. Establishment and characterization of immortalized human gingival keratinocyte cell lines. J Periodontal Res. 2008;43(6):604–14. doi:10.1111/j.1600-0765.2007.01019.x.

 PubMed Web of Science ®Google Scholar

Gröger S, et al. Effects of Porphyromonas gingivalis infection on human gingival epithelial barrier function in vitro. Eur J Oral Sci. 2010;118(6):582–9. doi:10.1111/j.1600-0722.2010.00782.x.

 PubMed Web of Science ®Google Scholar

Gröger S, et al. Influence of retinoic acid on human gingival epithelial barriers. J Periodontal Res. 2016;51(6):748–57. doi:10.1111/jre.12351.

 PubMed Web of Science ®Google Scholar

Kusumoto, Y, et al. Human gingival epithelial cells produce chemotactic factors interleukin-8 and monocyte chemoattractant protein-1 after stimulation with Porphyromonas gingivalis via toll-like receptor 2. J Periodontol. 2004;75(3):370–9. doi:10.1902/jop.2004.75.3.370.

 PubMed Web of Science ®Google Scholar

Umeda, JE, et al. Differential transcription of virulence genes in Aggregatibacter actinomycetemcomitans serotypes. J Oral Microbiol. 2013;5. doi:10.3402/jom.v5i0.21473.

 PubMed Web of Science ®Google Scholar

Savitri, IJ, et al. Irsogladine maleate inhibits Porphyromonas gingivalis-mediated expression of toll-like receptor 2 and interleukin-8 in human gingival epithelial cells. J Periodontal Res. 2015;50(4):486–93. doi:10.1111/jre.12231.

 PubMed Web of Science ®Google Scholar

Ouhara, K, et al. miR-584 expressed in human gingival epithelial cells is induced by Porphyromonas gingivalis stimulation and regulates interleukin-8 production via lactoferrin receptor. J Periodontol. 85, 2014;6:e198–204. doi:10.1902/jop.2013.130335.

 Google Scholar

Benso, B, et al. Malva sylvestris inhibits inflammatory response in oral human cells. An In Vitro Infection Model. PLoS One. 2015;10(10):e0140331. doi:10.1371/journal.pone.0140331.

 PubMed Web of Science ®Google Scholar

Castilho, et al. Rac1 is required for epithelial stem cell function during dermal and oral mucosal wound healing but not for tissue homeostasis in mice. 2010;5(5):e10503.

 Google Scholar

Glazer, CA, et al. The Role of MAGEA2 in head and neck cancer. Arch Otolaryngol Head Neck Surg. 2011;137(3):286–93. doi:10.1001/archoto.2011.2.

 PubMedGoogle Scholar

Bhan, S, et al. MAGEA4 induces growth in normal oral keratinocytes by inhibiting growth arrest and apoptosis. Oncol Rep. 2012;28(4):1498–502. doi:10.3892/or.2012.1934.

 PubMed Web of Science ®Google Scholar

Pellicioli, AC, et al. Laser phototherapy accelerates oral keratinocyte migration through the modulation of the mammalian target of rapamycin signaling pathway. J Biomed Opt. 2014;19(2):028002. doi:10.1117/1.JBO.19.2.028002.

 PubMed Web of Science ®Google Scholar

Martins, MD, et al. Epigenetic modifications of histones in periodontal disease. J Dent Res. 2016;95(2):215–22. doi:10.1177/0022034515611876.

 PubMed Web of Science ®Google Scholar

Dongari-Bagtzoglou A, Kashleva H. Development of a highly reproducible three-dimensional organotypic model of the oral mucosa. Nat Protoc. 2006;1(4):2012–8. doi:10.1038/nprot.2006.323.

 PubMed Web of Science ®Google Scholar

Dongari-Bagtzoglou, A, Kashleva, H. Candida albicans triggers interleukin-8 secretion by oral epithelial cells. Microb Pathog. 2003;34(4):169–77. doi:10.1016/S0882-4010(03)00004-4.

 PubMed Web of Science ®Google Scholar

Wöllert, T, et al. Human oral keratinocytes: a model system to analyze host-pathogen interactions. Methods Mol Biol. 2012;845:289–302. doi:10.1007/978-1-61779-539-8_19.

 PubMedGoogle Scholar

Volk, et al. Glutathione level and genotoxicity in human oral keratinocytes exposed to TEGDMA. J Biomed Mater Res B Appl Biomater. 2012;100( 2):391–9. doi:10.1002/jbm.b.31960.

 Google Scholar

Ramage, G, et al. Antifungal, cytotoxic, and immunomodulatory properties of tea tree oil and its derivative components: potential role in management of oral candidosis in cancer patients. Front Microbiol. 2012;3:220. doi:10.3389/fmicb.2012.00220.

 PubMed Web of Science ®Google Scholar

Millhouse, E, et al. Development of an in vitro periodontal biofilm model for assessing antimicrobial and host modulatory effects of bioactive molecules. BMC Oral Health. 2014;14:80. doi:10.1186/1472-6831-14-80.

 PubMed Web of Science ®Google Scholar

Wessels, M, et al. Genotoxic effects of camphorquinone and DMT on human oral and intestinal cells. Dent Mater. 2015;31(10):1159–68. doi:10.1016/j.dental.2015.06.007.

 PubMed Web of Science ®Google Scholar

Almela, T, Brook, IM, Moharamzadeh, K. Development of three-dimensional tissue engineered bone-oral mucosal composite models. J Mater Sci Mater Med. 2016;27:65. doi:10.1007/s10856-016-5676-7.

 PubMed Web of Science ®Google Scholar

Gilchrist, EP, et al. Establishment of a human polyclonal oral epithelial cell line. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000;90(3):340–7. doi:10.1067/moe.2000.107360.

 PubMed Web of Science ®Google Scholar

Kanno, F, Korostoff J, Volgina A, DiRienzo JM. Resistance of human periodontal ligament fibroblasts to the cytolethal distending toxin of Actinobacillus actinomycetemcomitans. J Peridontol. 2005;76(7):1189–201. doi:10.1902/jop.2005.76.7.1189.

 PubMed Web of Science ®Google Scholar

Feghali, K, Tanabe, S, Grenier, D. Soluble CD14 induces cytokine release by human oral epithelial cells. J Periodontal Res. 2011;46(1):147–52. doi:10.1111/j.1600-0765.2010.01311.x.

 PubMed Web of Science ®Google Scholar

Yee, M, et al. Porphyromonas gingivalis stimulates IL-6 and IL-8 secretion in GMSM-K, HSC-3 and H413 oral epithelial cells. Anaerobe. 2014;28:62–7. doi:10.1016/j.anaerobe.2014.05.011.

 PubMed Web of Science ®Google Scholar

Tabuchi, Y, et al. Development of oral epithelial cell line ROE2 with differentiation potential from transgenic rats harboring temperature-sensitive simian virus40 large T-antigen gene. Exp Anim. 1, 2014;63:31–44. doi:10.1538/expanim.63.31.

 Google Scholar

Tabuchi, Y, et al. Genes and gene networks involved in sodium fluoride-elicited cell death accompanying endoplasmic reticulum stress in oral epithelial cells. Int J Mol Sci. 2014;15(5):8959–78. doi:10.3390/ijms15058959.

 PubMed Web of Science ®Google Scholar

Jacobsen J, et al. TR146 cells grown on filters as a model for human buccal epithelium: I. Morphology, growth, barrier properties, and permeability. Int J Pharm. 1995;125:165–84. doi:10.1016/0378-5173(95)00109-V.

 Google Scholar

Portero A, Remuñán-López C, Nielsen HM. The potential of chitosan in enhancing peptide and protein absorption across the TR146 cell culture model-an in vitro model of the buccal epithelium. Pharm Res. 2002;19(2):169–74. doi:10.1023/A:1014220832384.

 PubMed Web of Science ®Google Scholar

Jacobsen J, et al. Filter-grown TR146 cells as an in vitro model of human buccal epithelial permeability. Eur J Oral Sci. 1999;107(2):138–46. doi:10.1046/j.0909-8836.1999.eos107210.x.

 PubMed Web of Science ®Google Scholar

Nielsen HM, et al. TR146 cells grown on filters as a model of human buccal epithelium: permeability of fluorescein isothiocyanate-labelled dextrans in the presence of sodium glycocholate. J Control Release. 1999;60;(2-3):223–33. doi:10.1016/S0168-3659(99)00081-4.

 Web of Science ®Google Scholar

Sander C, Nielsen HM, Jacobsen J. Buccal delivery of metformin: TR146 cell culture model evaluating the use of bioadhesive chitosan discs for drug permeability enhancement. Int J Pharm. 2013;458(2):254–61. doi:10.1016/j.ijpharm.2013.10.026.

 PubMedGoogle Scholar

Teubl BJ, et al. The oral cavity as a biological barrier system: design of an advanced buccal in vitro permeability model. Eur J Pharm Biopharm. 2013;84(2):386–93. doi:10.1016/j.ejpb.2012.10.021.

 PubMed Web of Science ®Google Scholar

Eirheim, HU, Bundgaard, C, Nielsen, HM. Evaluation of different toxicity assays applied to proliferating cells and to stratified epithelium in relation to permeability enhancement with glycocholate. Toxicol In Vitro. 2004;18(5):649–57. doi:10.1016/j.tiv.2004.02.003.

 PubMed Web of Science ®Google Scholar

Iyire A, Alayedi M and Mohammed AR. Pre-formulation and systematic evaluation of amino acid assisted permeability of insulin across in vitro buccal cell layers. Sci Rep. 2016;6:32498. doi:10.1038/srep32498.

 PubMed Web of Science ®Google Scholar

Ye, P, et al. CD24 regulated gene expression and distribution of tight junction proteins is associated with altered barrier function in oral epithelial monolayers. BMC Cell Biol. 2009;10:2. doi:10.1186/1471-2121-10-2.

 PubMed Web of Science ®Google Scholar

Ye P. Modulation of epithelial tight junctions by TGF-beta 3 in cultured oral epithelial cells. Aust Dent J. 2012;57(1):11–7. doi:10.1111/j.1834-7819.2011.01651.x.

 PubMed Web of Science ®Google Scholar

Meyle J, et al. Transepithelial electrical resistance and tight junctions of human gingival keratinocytes. J Periodontal Res. 1999;34(4):214–22. doi:10.1111/j.1600-0765.1999.tb02244.x.

 PubMed Web of Science ®Google Scholar

Damek-Poprawa M, et al. Cell junction remodeling in gingival tissue exposed to a microbial toxin. J Dent Res. 2013;92(6):518–23. doi:10.1177/0022034513486807.

 PubMed Web of Science ®Google Scholar

Acheampong, EA, et al. Molecular interactions of human immunodeficiency virus type 1 with primary human oral keratinocytes. J Virol. 2005;79(13):8440–53. doi:10.1128/JVI.79.13.8440-8453.2005.

 PubMed Web of Science ®Google Scholar

Belibasakis GN, et al. The expression of gingival epithelial junctions in response to subgingival biofilms. Virulence. 2015;6(7):704–9. doi:10.1080/21505594.2015.1081731.

 PubMed Web of Science ®Google Scholar

Aframian, DJ, et al. Absence of tight junction formation in an allogeneic graft cell line used for developing an engineered artificial salivary gland. Tissue Eng. 2002;8(5):871–8. doi:10.1089/10763270260424231.

 PubMedGoogle Scholar

Maria, OM, et al. Matrigel improves functional properties of human submandibular salivary gland cell line. Int J Biochem Cell Biol. 2011;43(4):622–31. doi:10.1016/j.biocel.2011.01.001.

 PubMed Web of Science ®Google Scholar

Bamforth, SD, et al. A dominant mutant of occludin disrupts tight junction structure and function. J Cell Sci. 1999;112(12):1879–88.

 PubMed Web of Science ®Google Scholar

Vogel, C, et al. Flt-1, but not Flk-1 mediates hyperpermeability through activation of the PI3-K/Akt pathway. J Cell Physiol. 2007;212(1):236–43. doi:10.1002/jcp.21022.

 PubMed Web of Science ®Google Scholar

Michikawa, H, Fujita-Yoshigaki, J, Sugiya, H. Enhancement of barrier function by overexpression of claudin-4 in tight junctions of submandibular gland cells. Cell Tissue Res. 2008;334(2):255–64. doi:10.1007/s00441-008-0689-2.

 PubMed Web of Science ®Google Scholar

Mitsui, R, et al. Maintenance of paracellular barrier function by insulin-like growth factor-I in submandibular gland cells. Arch Oral Biol. 2010;55(12):963–9. doi:10.1016/j.archoralbio.2010.07.023.

 PubMed Web of Science ®Google Scholar

He X, et al. A polarized salivary cell monolayer useful for studying transepithelial fluid movement in vitro. Pflugers Arch. 1998;435(3):375–81. doi:10.1007/s004240050526.

 PubMed Web of Science ®Google Scholar

Castro, R, et al. Ion transport in an immortalized rat submandibular cell line SMG-C6. Proc Soc Exp Biol Med. 2000;225(1):39–48. doi:10.1046/j.1525-1373.2000.22505.x.

 PubMed Web of Science ®Google Scholar

Mei M, et al. Claudin-3 is required for modulation of paracellular permeability by TNF-α through ERK1/2/slug signaling axis in submandibular gland. Cell Signal. 2015;27:10. doi:10.1016/j.cellsig.2015.07.002.

 Web of Science ®Google Scholar

Li J, et al. ZO-1 and -2 Are Required for TRPV1-Modulated Paracellular Permeability. J Dent Res. 2015;94(12):1748–56. doi:10.1177/0022034515609268.

 PubMed Web of Science ®Google Scholar

Cong X, et al. Occludin is required for TRPV1-modulated paracellular permeability in the submandibular gland. J Cell Sci. 2013;126(5):1109–21. doi:10.1242/jcs.111781.

 PubMed Web of Science ®Google Scholar

Cong, X, et al. Claudin-4 is required for modulation of paracellular permeability by muscarinic acetylcholine receptor in epithelial cells. J Cell Sci. 2015;128(12):2271–86. doi:10.1242/jcs.165878.

 PubMed Web of Science ®Google Scholar

Xiang, RL, et al. Claudin-4 is required for AMPK-modulated paracellular permeability in submandibular gland cells. J Mol Cell Biol. 2014;6(6):486–97. doi:10.1093/jmcb/mju048.

 PubMed Web of Science ®Google Scholar

Ding C, et al. Adiponectin increases secretion of rat submandibular gland via adiponectin receptors-mediated AMPK signaling. PLoS One. 2013;8(5):e63878. doi:10.1371/journal.pone.0063878.

 Web of Science ®Google Scholar

Quissell DO, et al. Development and characterization of SV40 immortalized rat parotid acinar cell lines. In Vitro Cell Dev Biol Anim. 1998;34(1):58–67. doi:10.1007/s11626-998-0054-5.

 PubMed Web of Science ®Google Scholar

Baker OJ, et al. Proinflammatory cytokines tumor necrosis factor-alpha and interferon-gamma alter tight junction structure and function in the rat parotid gland Par-C10 cell line. Am J Physiol Cell Physiol. 2008;295(5):C1191–201. doi:10.1152/ajpcell.00144.2008.

 PubMed Web of Science ®Google Scholar

Li, D, Mrsny, RJ. Oncogenic Raf-1 disrupts epithelial tight junctions via downregulation of occludin. J Cell Biol. 2000;148(4):791–800. doi:10.1083/jcb.148.4.791.

 PubMed Web of Science ®Google Scholar

Hamm-Alvarez, SF, et al. Etk/Bmx activation modulates barrier function in epithelial cells. Am J Physiol Cell Physiol. 2001;280(6):C1657–68. doi:10.1152/ajpcell.2001.280.6.C1657.

 PubMed Web of Science ®Google Scholar

Sato M, et al. Search for specific markers of neoplastic epithelial duct and myoepithelial cell lines established from human salivary gland and characterization of their growth in vitro. Cancer. 1984;54(12):2959–67. doi:10.1002/1097-0142(19841215)54:12%3c2959:: AID-CNCR2820541225%3e3.0.CO;2-5.

 PubMed Web of Science ®Google Scholar

Jang SI, et al. Establishment of functional acinar-like cultures from human salivary glands. J Dent Res. 2015;94(2):304–11. doi:10.1177/0022034514559251.

 PubMed Web of Science ®Google Scholar

Tran SD, et al. Primary culture of polarized human salivary epithelial cells for use in developing an artificial salivary gland. Tissue Eng. 2005;11(1–2):172–81. doi:10.1089/ten.2005.11.172.

 PubMedGoogle Scholar

Hegyesi O, et al. Evidence for active electrolyte transport by two-dimensional monolayers of human salivary epithelial cells. Tissue Eng Part C Methods. 2015;21(12):1226–36. doi:10.1089/ten.tec.2014.0614.

 PubMed Web of Science ®Google Scholar

Maria OM, et al. Matrigel improves functional properties of primary human salivary gland cells. Tissue Eng Part A. 2011a;17(9–10):1229–38. doi:10.1089/ten.tea.2010.0297.

 PubMed Web of Science ®Google Scholar

Szlávik V, et al. Differentiation of primary human submandibular gland cells cultured on basement membrane extract. Tissue Eng Part A. 2008;14(11):1915–26. doi:10.1089/ten.tea.2007.0208.

 PubMed Web of Science ®Google Scholar

Tran SD, et al. Re-engineering primary epithelial cells from rhesus monkey parotid glands for use in developing an artificial salivary gland. Tissue Eng. 2006;12(10):2939–48. doi:10.1089/ten.2006.12.2939.

 PubMedGoogle Scholar

Aframian DJ, et al. Characterization of murine autologous salivary gland graft cells: a model for use with an artificial salivary gland. Tissue Eng. 2004;10(5–6):914–20. doi:10.1089/1076327041348518.

 PubMedGoogle Scholar