The Effects of Lithium Chloride on Survival and Function of in Vitro Cultured Human Granulosa Cells

Purpose: To investigate the effect of lithium chloride (LiCl) on survival and functional activity of in vitro cultured human granulosa cells (GCs). Materials and Methods: GCs were isolated from follicular fluid of women undergoing IVF/ICSI. After RBCs removal, GCs were plated in a medium containing varying concentrations of LiCl including 1, 2 and 5mM for two weeks period. A culture without LiCl was taken as control group. To induce apoptosis, the medium of all cultures, on day 14 were replaced by serum free media and the culture was extended for further 24 hour. At the end of cultivation period, apoptosis was assessed by TUNEL and flowcytometry analysis. Furthermore to assess secretional function of the cells in various groups, in some cultures, FSH and androstondion hormones were added and after 48 hours, subsequent secretion of 17-b estradiol was determined by RIA analysis. Results: The result showed that in GCs treated with 2 and 5 mM LiCl, apoptosis was statistically decreased more than 50% compared to control group (P>0.05) and estradiol secretion increased more than 40% in the GCs treated with 1,2,5 mM LiCl compared to control group. Conclusion: These results suggest that lithium increase functional activity of GCs and can protect GCs against apoptosis.