Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells

• Robust induction of hPGCLCs from primed hiPSCs occurs via incipient mesoderm-like cells
• EpCAM and INTEGRINα6 are identified as markers for hPGCLC purification
• hPGCLCs avoid activation of a somatic program and undergo epigenetic reprogramming
• BLIMP1 stabilizes germline transcription and represses neuronal differentiation

SummaryMechanisms underlying human germ cell development are unclear, partly due to difficulties in studying human embryos and lack of suitable experimental systems. Here, we show that human induced pluripotent stem cells (hiPSCs) differentiate into incipient mesoderm-like cells (iMeLCs), which robustly generate human primordial germ cell-like cells (hPGCLCs) that can be purified using the surface markers EpCAM and INTEGRINα6. The transcriptomes of hPGCLCs and primordial germ cells (PGCs) isolated from non-human primates are similar, and although specification of hPGCLCs and mouse PGCs rely on similar signaling pathways, hPGCLC specification transcriptionally activates germline fate without transiently inducing eminent somatic programs. This includes genes important for naive pluripotency and repression of key epigenetic modifiers, concomitant with epigenetic reprogramming. Accordingly, BLIMP1, which represses somatic programs in mice, activates and stabilizes a germline transcriptional circuit and represses a default neuronal differentiation program. Together, these findings provide a foundation for understanding and reconstituting human germ cell development in vitro.

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