Effect of glial cell line-derived neurotrophic factor on mouse spermatogonial stem cells in vitro

Abstract

Objective To investigate the effect of glial cell line-derived neurotrophic factor(GDNF)on proliferation and differentiation of mouse spermatogonial stem cells in vitro.Methods The percoll discontinue density gradient centrifugation,followed by removing contaminated somatic cells through adhesion to plastic dishes,was used to purify the spermatogonial stem cells of Kunming mice.Mouse spermatogonial stem cells were confirmed by immunofluorescence and flow cytometry.The cells were divided into control groups and exprimental groups.The spermatogonial stem cells were cultured on sertoli cells.GDNF was added to medium of DMEM/F12.The growth of spermatogonial stem cells was determined by ELISA.The cell cycle of spermatogonial stem cells was determined by flow cytometry.Sperm was allied with ovum by intracytoplasmic sperm injection(ICSI).The chromosome figure and quantity were analysed after 3 days.Results A values of spermatogonial stem cells in experimental group were 0.362±0.031,0.448±0.028,0.502±0.062,0.556±0.045,0.621±0.072 in 3,6,9,12,15 days respectively;in control group,they were 0.365±0.045,0.377±0.053,0.402±0.071,0.432±0.019,0.461±0.037 respectively.Significant differences were noted between experimental group and controI group(P＜0.05).S period's quantities of DNA synthesizing were 20.86,26.34,31.23,37.54,28.02 in 3,6,9,12,15 days respectively,while they were 1.69,1.73.2.56,4.85,1.82 in 3,6,9,12,15 days in control group.Significant differences were noted between experimental group and control group(P＜0.05).The dual DNA could be found when sperm was integrated ovum after 3 days.Conclusions ICSI could identify spermatogial stem cells and translate into spermic cells in vitro.GDNF could improve the proliferation and differenciation of the spermatogonial stem cells in vitro. Key words: Spermatogonia; Glial cell line-derived neurotrophic factor; Cell culture; Mice