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

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  • Sivakamasundari Vijayakumar

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Roberta Sala

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Gugene Kang

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Angela Chen

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Michelle Ann Pablo

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Abidemi Ismail Adebayo

    (Stanford University School of Medicine
    Stanford University School of Medicine
    Stanford University)

  • Andrea Cipriano

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Jonas L. Fowler

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Danielle L. Gomes

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Lay Teng Ang

    (Stanford University School of Medicine)

  • Kyle M. Loh

    (Stanford University School of Medicine
    Stanford University School of Medicine)

  • Vittorio Sebastiano

    (Stanford University School of Medicine
    Stanford University School of Medicine)

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.

Suggested Citation

  • 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. Lo, 2023. "Monolayer platform to generate and purify primordial germ-like cells in vitro provides insights into human germline specification," Nature Communications, Nature, vol. 14(1), pages 1-19, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-41302-w
    DOI: 10.1038/s41467-023-41302-w
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