Bovine Follicular Cell Culture with Fatty Acids

Primary GC culture and bovine serum albumin (BSA)-FA conjugate preparations were done as described earlier (21 (link)). For testing the effects of actual follicular concentration of ALA (Sigma-Aldrich,L2376) and cis-9, trans-11CLA (Sigma-Aldrich,16413) the media were replaced every 48 h with supplemented α-MEM media containing different concentrations of ALA (20, 40, and 80 μM) or cis-9, trans-11 CLA (15, 30, and 60 μM) as BSA conjugates or non-conjugated BSA as vehicle control (Sigma-Aldrich A7030). All control wells received the same volume of BSA as the test groups with highest fatty acid concentration. The conditioned media collected on the 8th day of culture were stored at −20°C for steroid hormone estimation, while the remaining cells were lysed for RNA isolation.