Monolayer platform to generate and purify primordial germ-like cells in vitro provides insights into human germline specification

Sivakamasundari Vijayakumar, Roberta Sala, Gugene Kang, Angela Chen, Michelle Ann Pablo, Abidemi Ismail Adebayo, Andrea Cipriano, Jonas L. Fowler, Danielle L. Gomes, Lay Teng Ang, Kyle M. Loh () and Vittorio Sebastiano ()
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Sivakamasundari Vijayakumar: Stanford University School of Medicine
Roberta Sala: Stanford University School of Medicine
Gugene Kang: Stanford University School of Medicine
Angela Chen: Stanford University School of Medicine
Michelle Ann Pablo: Stanford University School of Medicine
Abidemi Ismail Adebayo: Stanford University School of Medicine
Andrea Cipriano: Stanford University School of Medicine
Jonas L. Fowler: Stanford University School of Medicine
Danielle L. Gomes: Stanford University School of Medicine
Lay Teng Ang: Stanford University School of Medicine
Kyle M. Loh: Stanford University School of Medicine
Vittorio Sebastiano: Stanford University School of Medicine

Nature Communications, 2023, vol. 14, issue 1, 1-19

Abstract: Abstract Generating primordial germ cell-like cells (PGCLCs) from human pluripotent stem cells (hPSCs) advances studies of human reproduction and development of infertility treatments, but often entails complex 3D aggregates. Here we develop a simplified, monolayer method to differentiate hPSCs into PGCs within 3.5 days. We use our simplified differentiation platform and single-cell RNA-sequencing to achieve further insights into PGCLC specification. Transient WNT activation for 12 h followed by WNT inhibition specified PGCLCs; by contrast, sustained WNT induced primitive streak. Thus, somatic cells (primitive streak) and PGCLCs are related—yet distinct—lineages segregated by temporally-dynamic signaling. Pluripotency factors including NANOG are continuously expressed during the transition from pluripotency to posterior epiblast to PGCs, thus bridging pluripotent and germline states. Finally, hPSC-derived PGCLCs can be easily purified by virtue of their CXCR4+PDGFRA-GARP- surface-marker profile and single-cell RNA-sequencing reveals that they harbor transcriptional similarities with fetal PGCs.

Date: 2023
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DOI: 10.1038/s41467-023-41302-w

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