Modulating the lipid profile of blastocyst cell membrane with DPPC multilamellar vesicles

Figures & data

Table 1. Effect of different forms of storage on the average diameter of MLVs at different concentrations. Results are presented as the mean (in nm) ± SEM, as previously published [32].

		Storage form
Treatment	DPPC conc.	No incubation	24 h incubation	48 h incubation
	(mmol/L)	DH ± SEM (nm)	DH ± SEM (nm)	DH ± SEM (nm)
Ultrapure Water	1.0	1146 ± 111.3^a A	998 ± 125.0^b A	860 ± 63.0^a A
	1.5	1073 ± 141.7^a A	684 ± 57.9^bcd B	716 ± 49.1^ab B
	2.0	621 ± 44.0^b A	358 ± 35.0^d B	388 ± 37.1^d B
Embryonic culture medium	1.0	414 ± 43.6^b B	1490 ± 202.5^a A	454 ± 32.1 ^cd B
	1.5	704 ± 88.7^b A	848 ± 129.0^bc A	604 ± 62.4^bc A
	2.0	500 ± 39.2^b AB	380 ± 34.9 ^cd B	539 ± 37.8^bcd A

Lowercase letters (column) indicate a difference (p < .05) between different concentrations for the same form of storage.

Uppercase letters (line) indicate a difference for the same concentration stored in different forms.

D_H: hydrodynamic diameter; SEM: standard error of the mean.

Table 2. Evaluation of the embryonic development rate (after 7 days of culture; D7) with or without MLVs added on the 5^th day of culture (D5).

Treatment	COCs	Embryonic development
		Cleavage (% mean ± SEM)	Early blastocysts (% mean ± SEM)	Blastocysts (% mean ± SEM)*	Expanded blastocysts (% mean ± SEM)	Hatched blastocysts (% mean ± SEM)	Total blastocysts (% mean ± SEM)*
Control	272	85 ± 2.2	6 ± 2.1	15 ± 2.5	20 ± 3.3	4 ± 1.8	47 ± 5.8
G1	273	88 ± 2.1	3 ± 1.5	15 ± 2.1	24 ± 1.3	2 ± 1.0	43 ± 2.8
G2	281	89 ± 2.6	3 ± 0.7	15 ± 2.9	21 ± 3.4	4 ± 1.5	44 ± 4.2
G3	270	85 ± 2.6	4 ± 1.3	13 ± 2.8	17 ± 3.5	3 ± 0.9	39 ± 2.9
G4	287	79 ± 3.5	2 ± 0.6	10 ± 1.7	16 ± 1.5	3 ± 1.3	33 ± 2.8
G5	274	84 ± 4.3	1 ± 0.8	14 ± 2.9	20 ± 3.7	0.9 ± 0.6	38 ± 4.8

*The data presented were transformed (log10) to a parametric distribution prior to statistical analysis.

COCs: cumulus-oocyte complexes; SEM: standard error of the mean.

Figure 1. MALDI-MS spectra in the positive ion mode for blastocysts co-cultured with MLVs (G1, G3, and G5) or without MLVs (control; CG).

Figure 2. PLS-DA for embryos cultured without MLVs (CG) or with different concentrations of MLVs (G1, G3, and G5). (A) PLS-DA 2 D and (B) PLS-DA 3 D score plots.

Figure 3. (A) PLS-DA 2 D for embryos cultured with MLVs (G1, G3, and G5). (B) PLS-DA 2 D for comparing embryos cultured without MLVs (CG) and with the highest concentration of MLVs (G5).

Table 3. Tentative identification of discriminant ions in the CG and MLV-treated group (G5) based on the MALDI-MS profile.

m/z	Lipid ion (carbons:unsaturations)^a	Reference
720.6	[PCe (32:0) + H]^+	Belaz et al. [27]
754.9	[PC (32:1) + Na]^+; [PC (34:4) + Na]^+	Belaz et al. [27]
759.0	[PC (34:2) + H]^+	Razza et al. [33]
779.1	[GP (42:4) + H]^+	*LMGP04010091
781.2	[PC (34:2)+Na]^+; [PC (36:5) + H]^+	Ferreira et al. [25]
797.3	[GP (44:5) + H]^+	*LMGP04010225

^aThe attribution of the ions in the table was performed according to the smallest mass difference observed in the MALDI-MS spectra and the distribution value of the compounds in the LIPID MAPS ID database (http://www.lipidmaps.org). Isomeric lipid species cannot be determined by high-resolution mass spectrometry. Assignments are based on previously recorded MS-MS data and literature reports and are thus tentative. *Possible assignment as indicated in the LIPID MAPS ID database (http://www.lipidmaps.org). PC: phosphatidylcholine; GP: glycerophospholipid; PCe: alkyl ether PC.

Figure 4. Boxplots showing the difference in the relative abundance of six significant ions in the CG compared with G5.

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Data availability statement

The authors confirm that the data that support the findings of this study are available from Marcelo Fábio Gouveia Nogueira, upon reasonable request.