【摘要】 目的探讨血清培养对大鼠精原干细胞生长周期的影响。方法采用percoll分离及差速贴壁法纯化精原干细胞,抗端粒酶逆转录酶免疫组化进行鉴定,用流式细胞仪对细胞生长周期进行判断。结果在有血清的培养基中培养的精原干细胞,其OD值逐渐升高(P<0.05);精原干细胞DNA合成期(S期)含量的增多。结论抗端粒酶逆转录酶免疫组化可作为鉴定精原干细胞提供较为充分的依据;血清能促进精原干细胞的体外增殖。更多还原

【Abstract】 Objective To investigate the effect of serumn on rat spermatogonial stem cells in vitro culture.Methods The percoll discontinue density gradient centrifugation,followed by removal of contaminating somatic cells through adhesion to plastic dishes,was used to purify the spermatogonial stem cells.The telomerase-reverse transcriptase(TERT) special antibody was used to identify the spermatogial stem cells.Purified cells were cultured in vitro for short term.and the MTT was used to observe the growth of the spermatogonial stem cells.The cell cycle of spermatogonial stem cells was determined by flow cytometry.Results The OD number of the spermatogial stem cells in the DMEM containing serum increased significantly.However,the OD number of the spermatogial stem cells in the DMEM without serum declined significantly(P<(0.05) vs blood serum group).The content of DNA synthesis phase of spermatogonial stem cells in the DMEM containing serum increased.However,the content of DNA synthesis phase of the spermatogial stem cells in the DMEM without serum declined.Conclusion The immunohistochemistry of TERT is enough to identify spermatogial stem cells,The blood serumn can improve the growth of the spermatogonial stem cells.更多还原