Characterization of migratory primordial germ cells in the aorta-gonad-mesonephros of a 4.5-week-old human embryo: a toolbox to evaluate in vitro early gametogenesis

AbstractSTUDY QUESTIONWhich set of antibodies can be used to identify migratory and early post-migratory human primordial germ cells (hPGCs)?STUDY FINDINGWe validated the specificity of 33 antibodies for 31 markers, including POU5F1, NANOG, PRDM1 and TFAP2C as specific markers of hPGCs at 4.5 weeks of development of Carnegie stage (CS12 –13), whereas KIT and SOX17 also marked the intra-aortic hematopoietic stem cell cluster in the aorta-gonad-mesonephros (AGM).WHAT IS KNOWN ALREADYThe dynamics of gene expression during germ cell development in mice is well characterized and this knowledge has proved crucial to allow the development of protocols for thein vitro derivation of functional gametes. Although there is a great interest in generating human gametesin vitro, it is still unclear which markers are expressed during the early stages of hPGC development and many studies use markers described in mouse to benchmark differentiation of human PGC-like cells (hPGCLCs). Early post-implantation development differs significantly between mice and humans, and so some germ cells markers, including SOX2, SOX17, IFITM3 and ITGA6 may not identify mPGCs and hPGCs equally well.STUDY DESIGN, SIZE, DURATIONThis immunofluorescence study investigated the expression of putative hPGC markers in the caudal part of a single human embryo at 4.5 weeks of development.PARTICIPANTS/MATERIALS, SETTING, METHODSWe have investigated by immunofluorescence the expression of a set of 33 antibodies for 31 mark...
Source: Molecular Human Reproduction - Category: Molecular Biology Source Type: research