title: Human amniotic fluid stem cells possess the potential to differentiate into primordial follicle oocytes in vitro.
abstract: Previous reports have demonstrated that embryonic stem cells were capable of differentiating into primordial germ cells through the formation of embryoid bodies that subsequently generated oocyte-like cells (OLCs). Such a process could facilitate studies of primordial follicle oocyte development in vitro and regenerative medicine. To investigate the pluripotency of human amniotic fluid stem cells (hAFSCs) and their ability to differentiate into germ cells, we isolated a CD117(+)/CD44(+) hAFSC line that showed fibroblastoid morphology and intrinsically expressed both stem cell markers (OCT4, NANOG, SOX2) and germ cell markers (DAZL, STELLA). To encourage differentiation into OLCs, the hAFSCs were first cultured in a medium supplemented with 5% porcine follicular fluid for 10 days. During the induction period, cell aggregates formed and syntheses of steroid hormones were detected; some OLCs and granulosa cell-like cells could be loosened from the surface of the culture dish. Cell aggregates were collected and replated in oocyte culture medium for an additional 7-10 days. OLCs ranging from 50 to 120 μm presenting zona pellucida were observed in cumulus-oocyte complexes; some OLCs developed spontaneously into multicell structures similar to preimplantation embryos. Approximately 2% of the hAFSCs differentiated to meiotic germ cells that expressed folliculogenesis- and oogenesis-associated markers. Although the in vitro maturation and fertilization potentials are as yet unproven, short-term (<25 days) and high-efficiency (>2%) derivation of OLCs from hAFSCs might provide a new approach to the study of human germ cell development in vitro.
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Yu
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Qiang
authors.forename: Qianhui
authors.initials: Q
authors.lastname: Wan
authors.forename: Mingming
authors.initials: M
authors.lastname: Qin
authors.forename: Shuai
authors.initials: S
authors.lastname: Chen
authors.forename: Huayan
authors.initials: H
authors.lastname: Wang
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D019070:Cell Lineage / Q000502:physiology; D003470:Culture Media / Q000494:pharmacology; D058732:Embryoid Bodies / Q000166:cytology; D005260:Female; D006801:Humans; D009865:Oocytes / Q000166:cytology*; D009866:Oogenesis / Q000502:physiology; D006080:Ovarian Follicle / Q000166:cytology*; D010312:Parthenogenesis / Q000502:physiology; D011247:Pregnancy; D013234:Stem Cells / Q000166:cytology*; D034421:Sus scrofa
chemical_list: D003470:Culture Media
keywords: differentiation; embryo; human reproduction; oocyte maturation; stem cells