Differentiation of rat iPS cells and ES cells into granulosa cell-like cells in vitro

Juan Zhang, Hui Li, Zhao Wu, XiaoJun Tan, Fengying Liu, Xianghong Huang and Xiaoling Fang

Acta Biochim Biophys Sin 2013, 45: 289–295; doi: 10.1093/abbs/gmt008

Department of Obstetrics and Gynecology, Second Xiangya Hospital, Central South University, Changsha 410083, China

Premature ovarian failure (POF) is an ovarian defect characterized by the premature depletion of ovarian follicles before 40 years of age, representing one major cause of female infertility. Stem cells provide the possibility of a potential treatment for POF. In this study, rat embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) were co-cultured with granulosa cells (GCs) to differentiate to GC-like cells. The level of estradiol (E2) analyzed by radioimmunoassay showed that the E2 concentration of the culture supernatant of co-cultured rat iPSCs and ESCs increased in a time-dependent manner, compared with the GCs group that has an opposite trend. The expression of follicle-stimulating hormone receptor (FSHR) was confirmed by immunostaining. These results indicated that rat iPSCs and ESCs were effectively induced to GC-like cells through indirect cell-to-cell contact. Real-time polymerase chain reaction was performed to analyze the expression level of marker genes in POF, including BMP15, FMR1, FSHR, INHA, AMH, NOBOX, FOXO3, EIF2B, FIGLA, and GDF9. The BMP15, FSHR, INHA, AMH, NOBOX, and GDF9 genes were significantly up-regulated in iPSCs and ESCs co-cultured with GCs in comparison with cells that were not co-cultured. Thus, here we demonstrated an available method to differentiate rat iPSCs and ESCs into GC-like cells in vitro for the possible cell therapy of POF.

Expression of FSHR in iPSCs and ESCs co-cultured with GCs

全文: http://abbs.oxfordjournals.org/content/45/4/289.full

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