人卵泡颗粒细胞的原代培养与鉴定

Primary Culture and Identification of Human Folliole Granulosa Cells

目的建立和完善人卵泡颗粒细胞的体外原代培养和鉴定方法。方法分离行辅助生殖助孕技术治疗的符合研究纳入标准的不孕患者的卵泡颗粒细胞,接种于DMEM培养基进行原代培养;以H-E染色和卵泡刺激素受体(FSHR)免疫组织化学染色对所培养的卵泡颗粒细胞进行鉴定。结果分离培养的人卵泡颗粒细胞存活率>90%;FSHR免疫组织化学鉴定细胞纯度达到95%以上;体外培养的人卵泡颗粒细胞在培养第2～5 d时处于分裂高峰。结论采用本研究所建立的方法可获得生长活性好、细胞纯度高及较稳定的人卵泡颗粒细胞,H-E染色和FSHR免疫组织化学染色可以简便快速地鉴定人卵泡颗粒细胞。

Objective To establish and improve the primary culture in vitro and identification methods of human folliole granulosa cells.Methods Primary granulosa cells were obtained from women who were meeting the inclusion criteria and undergoing ART.Folliole granulosa cells were incubated in the DMEM medium.H-E staining and FSHR immunohistochemistry methods were used to identify folliole granulosa cells.Results Over 90% of the cultured cells were folliole granulosa cells.Cell purity was above 95% by FSHR immunohistochemical identification.Cultured human folliole granulosa cells during the first 2～5d were to reach the splitting peak.Conclusions Good growth activity,cell purity and more stable human folliole granulosa cells could be obtained by the method used in this study.Human folliole granulosa cells could be simply and rapidly identified by H-E staining and FSHR immunohistochemistry methods.