Abstract
We recently reported a robust and defined culture system for the specification of human primordial germ cell-like cells (hPGCLCs) from human pluripotent stem cells (hPSCs), both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in vitro (Irie et al. Cell 160: 253–268, 2015). Similar attempts previously produced hPGCLCs from hPSCs at a very low efficiency, and the resulting cells were not fully characterized. A key step, which facilitated efficient hPGCLC specification from hPSCs, was the induction of a “competent” state for PGC fate via the medium containing a cocktail of four inhibitors. The competency of hPSCs can be maintained indefinitely and interchangeably with the conventional/low-competent hPSCs. Specification of hPGCLC occurs following sequential expression of key germ cell fate regulators, notably SOX17 and BLIMP1, as well as initiation of epigenetic resetting over 5 days. The hPGCLCs can be isolated using specific cell surface markers without the need for generating germ cell-specific reporter hPSC lines. This powerful method for the induction and isolation of hPGCLCs can be applied to both hESCs and iPSCs, which can be used for advances in human germ line biology.
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Acknowledgement
This work was supported by a Wellcome Trust Investigator Award, and by a BIRAX Grant. We thank Dr. Toshihiro Kobayashi, Dr. Roopsha Sengupta, and Dr. Carlos le Sage for critical reading of the manuscript. The institute is funded by a core grant from the Wellcome Trust (092096) and Cancer Research UK (C6946/A14492).
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Irie, N., Surani, M.A. (2017). Efficient Induction and Isolation of Human Primordial Germ Cell-Like Cells from Competent Human Pluripotent Stem Cells. In: Buszczak, M. (eds) Germline Stem Cells. Methods in Molecular Biology, vol 1463. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4017-2_16
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DOI: https://doi.org/10.1007/978-1-4939-4017-2_16
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