解剖学杂志(社会科学版)2019，Vol.42Issue(2)：116-120,5.DOI:10.3969/j.issn.1001-1633.2019.02.002
人卵巢颗粒细胞分离培养的优化及生物学特性
Culture technique optimization and biological characteristics of human ovarian granulosa cells
摘要
Abstract
Objective:To establish the methods of isolating and culturing human ovarian granulosa cells(hGCs)and observe biological characteristics.Methods:hGCs were harvested during oocyte retrieval.Then they were divided into control group(DMEM medium)and experimental group(DMEM medium:follicular fluid=1:1).The characteristic of hGCs and secretion of estrogen were observed.The expression levels of Nanog,Oct4 and Sox2 were detected by RT-qPCR.CD44,CD73 and CD45 were measured via flow cytometry(FCM).Results:The hGCs grew well in vitro after 24 h,and cells were connected by pseudopodia.Apoptosis began in the cells of control group after 14 d,while cells of experimental group began to degenerate until 21 d.Compared with control group,the secretion of estrogen in the experimental group increased significantly.It reached the peak at 9 d and significantly declined at 21 d.The results from RT-qPCR indicated that the expression levels of Oct4 and Nanog of experimental group increased significantly at 7 d.FCM analysis showed that CD44 and CD73 were positive and CD45 was negative after 7 d in vitro culture.Conclusion:This method is simple and effective to obtain a large number of hGCs.Compared with the culture medium,adding follicular fluid is beneficial to the growth of hGCs,and increase the culture time.Meanwhile,hGCs can secrete estrogen and has certain stem cell characteristics.关键词
人卵巢颗粒细胞/细胞培养/细胞沉淀法/生物学特性/干细胞Key words
human granulosa cell/cell culture/cell precipitation/biological characteristic/stem cell引用本文复制引用
许素铭,王耀琴,朱鹏飞,李宵,王春芳,武学清..人卵巢颗粒细胞分离培养的优化及生物学特性[J].解剖学杂志(社会科学版),2019,42(2):116-120,5.基金项目
国家重点研发计划(2018YFC1002103) （2018YFC1002103）
山西省回国留学人员科研资助项目(2012-100) （2012-100）
