ABSTRACT
Objective To improve the current method of spermatogonial cells transplantation, and make the new method become more operative and easy to carry out. Methods The spermatogonial cells in rC57BL6/tg14 (act-EGFO-Osb Y01) mice that expresse GFP protein were collected as the donor cells, and by using a modified syringe, and then were injected into the seminiferous tubules of pretreated wild type GFP-null mice through testicular efferent duct. The transplantation outcome was evaluated by trypan blue stainting and fluorescent microscopic examination and pathohistological analysis of transplanted testis. Results The transplanted testis of recipient mice showed green fluorescence signal, and the signal of seminiferous tubules was found significantly higher than that of the surrounding tissues. The transplanted GFP-positive cells generated colonies and spermatogenesis but pretreated wild type GFP-null mice were not found. Conclusion The expressed GFP protein spermatogonial cells in rC57BL6/tg14 mice were successfully transplanted in the wild type GFP-null recipient mice, fourthermore the improved transplantation method simplified the triditional one and achieved the same transplantation results.
ABSTRACT
Aim To investigate the protective effects of ginsenosides ( GS) on reactive oxygen species-induced oxidative damage in mouse spermatogonial cells. Meth-ods Mouse spermatogonial cell oxidative stress model was established and the attenuating effects of ginsen-osides on germ cell oxidative damage were evaluated by determination of cell viability,malondialdehyde( MDA) formation,superoxide dismutase ( SOD) activity and glutathione ( GSH) level. Results The exposure to hypoxanthine/xanthine oxidase ( HX/XO) induced an elevation in MDA,while a decrease in germ cell viability,SOD activity and GSH level. However,supplementation with GS ( 10 mg?L -1) restored HX/XO-induced decrease in cell viability,SOD activity and GSH level and HX/XO-induced increase in MDA formation. Conclusion GS may exert antioxidant activity to attenuate reactive oxygen species-induced oxidative damage in mouse spermatogonial cells.
ABSTRACT
Objective To study isolation and identification and culture of rat type A spermatogonial cells in vitro.Methods Percoll discontinue density gradient centrifugation combined with different speeds of different cells adhering to dish was used to purify the type A spermatogonial cells.The c-kit and TERT special antibodies were used to identify the type A spermatogonial cells.The purified cells were cultured in vitro. Results 0.614?10~6 cells per testis finally were obtained and the percentage of viable cells was 92.1% by trypan blue dye exclusion test.The percentage of type A spermatogonial cells expressing c-kit and TERT were 91.7?1.2% and 90.8?1.0% respectively.Type A spermatogonial cells could proliferate and self-renew in the DMEM containing 10% NBS.Conclusion Percoll discontinue density gradient centrifugation combined with different speeds of different cells adhering to dish is an efficiency method for isolation of rat type A spermatogonial cells.The purified cells are type A spermatogonial cells by identification of the immunohistochemistry of c-kit and TERT antibodies.Type A spermatogonial cells can proliferate and self-renew in vitro.