The generation of research-quality, clinically relevant cell types in vitro from human pluripotent stem cells requires a detailed understanding of the equivalent human cell types. Here we analysed 134 human embryonic and fetal samples from 6 to 20 developmental weeks and identified the stages at which cKIT+ primordial germ cells (PGCs), the precursors of gametes, undergo whole-genome epigenetic reprogramming with global depletion of 5mC, H3K27me3 and H2A.Z, and the time at which imprint erasure is initiated and 5hmC is present. Using five alternative in vitro differentiation strategies combined with single-cell microfluidic analysis and a bona fide human cKIT+ PGC signature, we show the stage of cKIT+ PGC formation in the first 16 days of differentiation. Taken together, our study creates a resource of human germ line ontogeny that is essential for future studies aimed at in vitro differentiation and unveiling the mechanisms necessary to pass human DNA from one generation to the next.
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The authors would like to thank the UCLA Translational Pathology Core Laboratory and the UCLA Gene and Cellular Core Laboratory for some of the gonadal samples used in this study. We also thank J. Hargan-Calvopina, M. Oliveros-Etter and S. Diaz-Perez for critical reading of the manuscript, F. Codrea and J. Scholes for FACS and S. Peckman from the Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research for critical assistance with human subject and embryonic stem cell review. This work was supported primarily by fund number 1R01HD058047 from the Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD; ATC), as well as the Iris Cantor-UCLA Women’s Health Pilot Project (ATC) and 1P01GM081621 from NIGMS. The Laboratory of Developmental Biology, University of Washington, Seattle is supported by NIH Award Number 5R24HD000836 from the NICHD. Human fetal tissue requests can be made to: firstname.lastname@example.org.
The authors declare no competing financial interests.
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Gkountela, S., Li, Z., Vincent, J. et al. The ontogeny of cKIT+ human primordial germ cells proves to be a resource for human germ line reprogramming, imprint erasure and in vitro differentiation. Nat Cell Biol 15, 113–122 (2013). https://doi.org/10.1038/ncb2638
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