title: BMP15 gene is activated during human amniotic fluid stem cell differentiation into oocyte-like cells.
abstract: The generation of oocyte-like cells (OLCs) from stem cell differentiation in vitro provides an optimal approach for studying the mechanism of oocyte development and maturation. The aim of this study was to investigate the activation of bone morphogenetic protein 15 gene (BMP15) during the differentiation of human amniotic fluid stem cells (hAFSCs) into OLCs. After 15 days of differentiation, OLCs with a diameter of 50-60 μm and zona pellucida (ZP)-like morphology were observed. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed the BMP15 was activated from approximately day 10 of differentiating hAFSCs and thereafter. The reporter construct pBMP15-enhanced green fluorescent protein (EGFP) was transiently transfected into the differentiated hAFSCs and the EGFP expression driven by the BMP15 promoter was positive in the OLCs. Moreover, RT-PCR analysis showed that the oocyte-specific markers including ZP1, ZP2, ZP3, and c-kit were expressed in the differentiated hAFSCs, and the immunofluorescence assay confirmed that the ZP2 was detected in the OLCs. Quantitative RT-PCR revealed that ZP2 and ZP3 were significantly elevated in the differentiated hAFSCs. Further, in the OLCs derived from hAFSCs, the BMP15 promoter directing the EGFP reporter was colocalized with ZP2. Together, these results illustrated that the BMP15 could be used as an oogenesis marker to track hAFSCs differentiation into the OLCs.
authors.affiliation: Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Cheng
authors.forename: Shuai
authors.initials: S
authors.lastname: Chen
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Yu
authors.forename: Pengsheng
authors.initials: P
authors.lastname: Zheng
authors.forename: Huayan
authors.initials: H
authors.lastname: Wang
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D055430:Bone Morphogenetic Protein 15 / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics*; D002470:Cell Survival; D003470:Culture Media / Q000378:metabolism; D005260:Female; D015571:Follicular Fluid / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics; D005819:Genetic Markers / Q000235:genetics; D006801:Humans; D009865:Oocytes / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: C525901:BMP15 protein, human; D055430:Bone Morphogenetic Protein 15; D003470:Culture Media; D005819:Genetic Markers