Yanfei Wen1,2, Wen He1, Manbo Jiang1,3, Minhui Zeng1,4, Liuhong Cai1. 1. Center for Reproductive Medicine, The Third Affiliated Hospital, Sun Yat-sen University, 6 East Longkou Road, Guangzhou, China. 2. Center for Reproductive Medicine, Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-sen University, Jiangmen, China. 3. Department of Reproductive Medicine, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, China. 4. Memorial hospital of Sun Yat-sen University, Guangzhou, China.
Abstract
AIM: We proposed a two-step protocol for deriving cells expressing markers of female germ cells (FGCs) from premature ovarian failure patient-specific induced pluripotent stem cells (POF-iPSCs). MATERIAL & METHODS: We cultured POF-iPSCs in suspension and pretreated them with TGFβ-1 (1 ng/ml) for 2 days and continued with both TGFβ-1 and BMP4 (50 ng/ml) for 5 more days. Then changed to media containing retinoic acid (1 μM) and 5% follicular fluid for another 7 days. Expression of markers of different stages of FGCs were detected. RESULTS: c-KIT, STELLA/DPPA3, VASA/DDX4, SCP3, GDF9 and ZP3 were positively detected and statistically significant different when compared with control groups. CONCLUSION: Our in vitro system was beneficial for POF-iPSCs differentiated cells to express STELLA, VASA and SCP3, which were the markers of meiosis initiation of FGCs.
AIM: We proposed a two-step protocol for deriving cells expressing markers of female germ cells (FGCs) from premature ovarian failurepatient-specific induced pluripotent stem cells (POF-iPSCs). MATERIAL & METHODS: We cultured POF-iPSCs in suspension and pretreated them with TGFβ-1 (1 ng/ml) for 2 days and continued with both TGFβ-1 and BMP4 (50 ng/ml) for 5 more days. Then changed to media containing retinoic acid (1 μM) and 5% follicular fluid for another 7 days. Expression of markers of different stages of FGCs were detected. RESULTS:c-KIT, STELLA/DPPA3, VASA/DDX4, SCP3, GDF9 and ZP3 were positively detected and statistically significant different when compared with control groups. CONCLUSION: Our in vitro system was beneficial for POF-iPSCs differentiated cells to express STELLA, VASA and SCP3, which were the markers of meiosis initiation of FGCs.