Derivation of embryonic germ cells and male gametes from embryonic stem cells

Abstract

Egg and sperm cells (gametes) of the mouse are derived from a founder population of primordial germ cells that are set aside early in embryogenesis. Primordial germ cells arise from the proximal epiblast, a region of the early mouse embryo that also contributes to the first blood lineages of the embryonic yolk sac1. Embryonic stem cells differentiate in vitro into cystic structures called embryoid bodies consisting of tissue lineages typical of the early mouse embryo2,3. Because embryoid bodies sustain blood development, we reasoned that they might also support primordial germ cell formation. Here we isolate primordial germ cells from embryoid bodies, and derive continuously growing lines of embryonic germ cells. Embryonic germ cells show erasure of the methylation markers (imprints) of the Igf2r and H19 genes, a property characteristic of the germ lineage. We show that embryoid bodies support maturation of the primordial germ cells into haploid male gametes, which when injected into oocytes restore the somatic diploid chromosome complement and develop into blastocysts. Our ability to derive germ cells from embryonic stem cells provides an accessible in vitro model system for studies of germline epigenetic modification and mammalian gametogenesis.

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Figure 1: Expression of germ-cell-specific genes during EB development.
Figure 2: Development of primordial germ cells in the differentiating EB.
Figure 3: RT–PCR detection of germ-cell-specific genes in SSEA1+ cells isolated from developing EBs.
Figure 4: EBs support differentiation of haploid male germ cells that support fertilization of oocytes.

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Acknowledgements

The authors thank S. L. Opitz, N. Watson and E. Dikovskaia for technical support; K. Hochedlinger and T. Holm for assistance with intracytoplasmic oocyte injection; and W. Lensch for critical revisions of the manuscript. This work was supported by grants from the National Institutes of Health, the National Science Foundation Biotechnology Process Engineering Center, and the Dutch Cancer Society ‘Koningin Wilhelmina fonds’. G.Q.D. is a Birnbaum Scholar of the Leukemia and Lymphoma Society of America. J.G. was sponsored by the Human Frontiers Science Foundation. K.E. is a Junior Fellow in the Harvard Society of Fellows.

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Correspondence to George Q. Daley.

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Geijsen, N., Horoschak, M., Kim, K. et al. Derivation of embryonic germ cells and male gametes from embryonic stem cells. Nature 427, 148–154 (2004). https://doi.org/10.1038/nature02247

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