Human Primordial Germ Cells Are Specified from Lineage-Primed Progenitors

Di Chen; Na Sun; Lei Hou; Rachel Kim; Jared Faith; Marianna Aslanyan; Yu Tao; Yi Zheng; Jianping Fu; Wanlu Liu; Manolis Kellis; Amander Clark

doi:10.1016/j.celrep.2019.11.083

Human Primordial Germ Cells Are Specified from Lineage-Primed Progenitors

Cell Rep. 2019 Dec 24;29(13):4568-4582.e5. doi: 10.1016/j.celrep.2019.11.083.

Authors

Di Chen¹, Na Sun², Lei Hou², Rachel Kim³, Jared Faith¹, Marianna Aslanyan¹, Yu Tao¹, Yi Zheng⁴, Jianping Fu⁵, Wanlu Liu⁶, Manolis Kellis⁷, Amander Clark⁸

Affiliations

¹ Department of Molecular Cell and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² MIT Computer Science and Artificial Intelligence Laboratory, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
³ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁴ Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.
⁵ Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI 48109, USA; Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.
⁶ Zhejiang University-University of Edinburgh Institute, Zhejiang University School of Medicine, 310058 Hangzhou, PR China.
⁷ MIT Computer Science and Artificial Intelligence Laboratory, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: manoli@mit.edu.
⁸ Department of Molecular Cell and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA; Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: clarka@ucla.edu.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Amnion / cytology
Animals
Cell Line
Cell Lineage*
Embryo, Mammalian / cytology
Embryonic Development
Gastrulation
Germ Cells / cytology*
Human Embryonic Stem Cells / cytology
Humans
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / metabolism
Mice
Models, Biological
Mutation / genetics
Primitive Streak / cytology
SOXF Transcription Factors / metabolism
Stem Cells / cytology*
Transcription Factor AP-2 / metabolism

Substances

SOX17 protein, human
SOXF Transcription Factors
TFAP2C protein, human
Transcription Factor AP-2

Abstract

Publication types

MeSH terms

Substances

Grants and funding