Docosahexaenoic acid (DHA) effects on proliferation and steroidogenesis of bovine granulosa cells

Table 2 Fatty acid composition from total lipids of bovine granulosa cells after 15 h treatment with DHA

Fatty acids (mole % of total fatty acids)	Control ^a	DHA1 μM	DHA 10 μM	DHA 50 μM
Fatty acids (mole % of total fatty acids)	n = 5	n = 4	n = 4	n = 3
Saturates
16:0	31.50 ± 7.48	40.16 ± 9.38	44.12 ± 11.60	48.45 ± 7.44
18:0	10.68 ± 2.25	13.36 ± 3.03	15.04 ± 3.89	16.24 ± 2.60
Monounsaturates
16:1 n-7	2.32 ± 0.55	3.03 ± 0.72	3.33 ± 0.91	3.52 ± 0.68
18:1 n-7	8.93 ± 2.17	11.51 ± 2.85	12.36 ± 3.72	12.22 ± 3.01
18:1 n-9 cis	29.99 ± 7.18	38.91 ± 9.08	42.49 ± 11.58	43.03 ± 7.86
n-6 PUFA
18:2 n-6 cis	6.05 ± 1.29	7.87 ± 1.81	8.59 ± 2.33	8.41 ± 2.07
20:4 n-6	8.85 ± 2.12	11.87 ± 3.08	13.02 ± 3.99	11.24 ± 4.10
n-3 PUFA
18:3 n-3	0.51 ± 0.12	0.66 ± 0.15	0.74 ± 0.18	0.67 ± 0.17
20:5 n-3	0.98 ± 0.25	1.34 ± 0.31	1.60 ± 0.38	1.37 ± 0.42
22:5 n-3	7.03 ± 1.72	9.33 ± 2.28	10.04 ± 2.90	7.79 ± 3.31
22:6 n-3 (DHA)	0.84 ± 0.16	2.17 ± 0.04 *	10.27 ± 0.11 *	23.05 ± 6.58 *

Granulosa cells (GC) (2.5 × 10⁵ viable cells/250 μL media/well) were cultured into 48-well dishes in modified McCoy’s 5A media in the presence or absence of DHA (1, 10 or 50 μM). After 15 h culture, supernatants were removed and cells with the same treatment (12 wells) were pooled and stored under nitrogen gas at −80 °C until the total lipid extraction. The fatty acid analysis was performed by gas chromatography by the Plateforme de Lipidomique Fonctionnelle (INSA, Villeurbanne, France). The relative amount of each fatty acid is expressed as the area of each fatty acid peak, relative to the total area for all fatty acids (mole%). The results are expressed as means ± SEM (n = 4 per treatment). ^a Control corresponds to the serum-free modified McCoy’s 5A medium supplemented with DMSO (1:2000) as carrier solvent of DHA. The prepared mediums with the different concentrations of DHA were all adjusted for DMSO at 1:2000. ^* indicates a significant difference with control condition (Kruskal-Wallis test with Tukey’s multiple comparison test as post-hoc test, p < 0.05)

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