@article{d31ccf80f2fd44d3a243251694c9ac64,
title = "Human Primordial Germ Cells Are Specified from Lineage-Primed Progenitors",
abstract = "In vitro gametogenesis is the process of making germline cells from human pluripotent stem cells. The foundation of this model is the quality of the first progenitors called primordial germ cells (PGCs), which in vivo are specified during the peri-implantation window of human development. Here, we show that human PGC (hPGC) specification begins at day 12 post-fertilization. Using single-cell RNA sequencing of hPGC-like cells (hPGCLCs) differentiated from pluripotent stem cells, we discovered that hPGCLC specification involves resetting pluripotency toward a transitional state with shared characteristics between naive and primed pluripotency, followed by differentiation into lineage-primed TFAP2A+ progenitors. Applying the germline trajectory to TFAP2C mutants reveals that TFAP2C functions in the TFAP2A+ progenitors upstream of PRDM1 to regulate the expression of SOX17. This serves to protect hPGCLCs from crossing the Weismann's barrier to adopt somatic cell fates and, therefore, is an essential mechanism for successfully initiating in vitro gametogenesis.",
keywords = "TFAP2A, TFAP2C, germ cells, pluripotency, single cell RNA-sequencing, stem cells",
author = "Di Chen and Na Sun and Lei Hou and Rachel Kim and Jared Faith and Marianna Aslanyan and Yu Tao and Yi Zheng and Jianping Fu and Wanlu Liu and Manolis Kellis and Amander Clark",
note = "Funding Information: The authors would like to thank Felicia Codrea, Jessica Scholes, and Jeffery Calimlim for FACS, Jinghua Tang for banking and culturing of the UCLA hESC lines, and Steven Peckman from the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research (BSCRC) for critical assistance with human subject and embryonic stem cell research oversight review. The authors would also like to thank Professor Katja-Layland from the Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany, and the Department of Women's Health, Research Institute for Women's Health, Tubingen, Germany for the early 4- to 5-week de-identified human genital ridge samples. This work is supported by funds from an anonymous donor, as well as NIH/NICHD R01 HD079546 (A.C.), NIH NCATS UCLA CTSI grant number UL1TR0001881, and the UCLA David Geffen School of Medicine Regenerative Medicine Theme Award (A.C.). All human pre-implantation embryo and human embryo attachment culture studies were performed using funds from the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research Innovation Award. No NIH funds were used for research with human pre-implantation embryos. Human fetal tissue research is supported by a grant to Ian Glass at the University of Washington Birth Defects laboratory, 5R24HD000836-53. Human conceptus tissue requests can be made to bdrl@u.washington.edu. D.C. N.S. L.H. and A.C. designed the experiments. D.C. M.A. and J.F. conducted the experiments. N.S. and L.H. analyzed the scRNA-seq data; W.L. analyzed the ChIP-seq data. Y.Z. and J.F. conducted the scRNA-seq of modeled human amnion cells. D.C. N.S. L.H. M.K. and A.C. interpreted the data, and D.C. N.S. L.H. and A.C. wrote the manuscript. The authors declare no competing interests. Funding Information: The authors would like to thank Felicia Codrea, Jessica Scholes, and Jeffery Calimlim for FACS, Jinghua Tang for banking and culturing of the UCLA hESC lines, and Steven Peckman from the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research (BSCRC) for critical assistance with human subject and embryonic stem cell research oversight review. The authors would also like to thank Professor Katja-Layland from the Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany, and the Department of Women{\textquoteright}s Health, Research Institute for Women{\textquoteright}s Health, Tubingen, Germany for the early 4- to 5-week de-identified human genital ridge samples. This work is supported by funds from an anonymous donor, as well as NIH/NICHD R01 HD079546 (A.C.), NIH NCATS UCLA CTSI grant number UL1TR0001881 , and the UCLA David Geffen School of Medicine Regenerative Medicine Theme Award (A.C.). All human pre-implantation embryo and human embryo attachment culture studies were performed using funds from the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research Innovation Award . No NIH funds were used for research with human pre-implantation embryos. Human fetal tissue research is supported by a grant to Ian Glass at the University of Washington Birth Defects laboratory, 5R24HD000836-53 . Human conceptus tissue requests can be made to bdrl@u.washington.edu . Publisher Copyright: {\textcopyright} 2019 The Author(s)",
year = "2019",
month = dec,
day = "24",
doi = "10.1016/j.celrep.2019.11.083",
language = "English (US)",
volume = "29",
pages = "4568--4582.e5",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "13",
}