Effects of Wharton’s jelly-derived mesenchymal stem cells on neonatal neutrophils

Figures & data

Figure 1 Effects of WJ-MSCs on neutrophil apoptosis.

Notes: Adult and neonatal neutrophils were incubated with WJ-MSCs at varying ratios for 24 hours and then analyzed for apoptosis by flow cytometry using BD FACSArray quadrant and two-dimensional histogram statistics based on relative fluorescence. Each bar represents the mean ± standard error (n=3–8). ⁺Significantly different (P<0.05) from adult; *significantly different (P<0.05) from untreated control (0).

Abbreviation: MSCs, mesenchymal stem cells; WJ-MSCs, MSCs derived from Wharton’s jelly.

Figure 2 Expression of VEGF, catalase, HO-1, and NOX-1 in neutrophils exposed to WJ-MSCs.

Notes: Adult and neonatal neutrophils were incubated with WJ-MSCs (WJ-MSCs + PMN, 1:20) or medium control (PMN) for 4 hours, in the presence or absence of neutralizing antibodies to ICAM-1 or IL-6 (1 μg/mL). Total RNA was extracted, complementary DNA generated, and gene expression quantified by real-time polymerase chain reaction. Results were normalized to β-actin expression. Each bar represents the mean ± standard error (n=5–7). *Significantly different (P<0.05) from control; ⁺significantly different (P<0.05) from MSCs + PMN.

Abbreviations: NOX-1, NADPH oxidase-1; HO-1, heme oxygenase-1; VEGF, vascular endothelial growth factor; MSCs, mesenchymal stem cells; WJ-MSCs, MSCs derived from Wharton’s jelly; PMN, polymorphonuclear neutrophils; IL-6, interleukin-6; ICAM-1, intercellular adhesion molecule-1.

Figure 3 Expression of TLR-4 and COX-2 in neutrophils and WJ-MSCs.

Notes: (A) Adult and neonatal neutrophils were incubated in medium control (PMN), LPS (1 μg/mL) and/or WJ-MSCs (WJ-MSCs to neutrophils, 1:20) (B) WJ-MSCs were incubated in medium control (MSCs), with LPS (1 μg/mL), and/or adult or neonatal neutrophils (WJ-MSCs to neutrophils, 1:20). Total RNA was extracted from neutrophils (A) or WJ-MSCs (B), complementary DNA generated, and gene expression quantified by real-time polymerase chain reaction. Results were normalized to β-actin expression. Each bar represents the mean ± standard error (n=5–7). *Significantly different (P<0.05) from PMN control; ⁺significantly different (P<0.05) from PMN + LPS (A) or MSCs + LPS (B).

Abbreviations: COX-2, cyclooxygenase-2; MSCs, mesenchymal stem cells; WJ-MSCs, MSCs derived from Wharton’s jelly; PMN, polymorphonuclear neutrophils; LPS, lipopolysaccharide; TLR4, Toll-like receptor-4.

Table 1 Production of inflammatory mediators by neutrophils and WJ-MSCs

	Neutrophils, mean (95% CI)	WJ-MSCs mean (95% CI)	Neutrophil + WJ-MSCs (expected) mean (95% CI)	Neutrophil + WJ-MSCs (observed) mean (95% CI)
Adult
MIP-1β (pg/mL)	123 (42, 203)	17 (14, 20)	140 (56, 223)	72 (32, 112)
IL-8 (ng/mL)	12 (5, 20)	35 (11, 57)	47 (16, 77)	9 (3, 16)**
ICAM-1 (pg/mL)	89 (43, 134)	144 (101, 187)	233 (144, 321)	205 (142, 268)
Neonatal
MIP-1β (pg/mL)	66 (14, 119)	21 (15, 26)	87 (29, 145)	126 (52, 200)
IL-8 (ng/mL)	3 (1, 5)	24 (1, 48)	27 (2, 53)	29 (7, 50)
ICAM-1 (pg/mL)	102 (46, 157)	143 (50, 236)	230 (96, 393)	184 (84, 284)

Note:

** Significantly different (P<0.05) from neutrophil + WJ-MSCs (expected).

Abbreviations: CI, confidence interval; MSCs, mesenchymal stem cells; WJ-MSCs, MSCs derived from Wharton’s jelly; IL, interleukin; ICAM-1, intercellular adhesion molecule-1; MIP-1β, macrophage inflammatory protein-1β.