Effects of ascorbic acid and NAC on survival of BFGCs in vitro

In order to investigate effects of ascorbic acid(AA), N-acetyl-L-cysteine (NAC) antioxidants on survival of buffalo follicular granulosa cells (BFGCs) after serum withdrawal in vitro, the experiments were conducted with or without adding one of the antioxidants ( AA, NAC) to the FBS-deprived medium. After BFGCs were cultivated in a DMEM/F12 medium with fetal bovine serum (FBS) in vitro, FBS was withdrawn. After six hours the BFGCs growth, apoptosis-like changes, and BFGCs survival number was examined by multifunctional inversion microscope and MTT method,respectively. The results showed that the BFGCs apoptosis-like changes such as cell shrinkage, condensed nuclei, rounded shape, droping-off wall existed in all control groups and experimental groups treated with AA and NAC. In AA groups, the least apoptosis-like BFGCs were found in 0.5 and 1.0mmol/L groups, and no living cells were found in 150mmol/L group. The living BFGCs number was significantly higher in 0.5mmol/L group than in control group and in other test groups(P0.05) except 1mmol/L group. In the NAC groups, the least apoptosis-like BFGCs were found in 5 mmol/Lgroup, the number of apoptosis-like BFGCs increased gra- dually in other 5 test groups with the concentrations of NAC. The living BFGCs numberwas significantly higher in 5mmol/L group than that in other groups (P0.05). These results suggested that a lower concentration of AA or NAC can accelerate BFGCs survival and a higher concentration of that can suppressBFGCs survival in vitro after FBS was removed from medium.