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

机译：从胚胎干细胞衍生出胚胎生殖细胞和雄配子

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摘要

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 sac. Embryonic stem cells differentiate in vitro into cystic structures called embryoid bodies consisting of tissue lineages typical of the early mouse embryo. 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 embryold 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.
机译：小鼠的卵子和精子细胞（配子）来源于原始生殖细胞的原始种群，这些原始细胞在胚胎发生的早期就被搁置了。原始生殖细胞来自近端上皮细胞，这是早期小鼠胚胎的一个区域，该区域也有助于卵黄囊的最初血统。胚胎干细胞在体外分化成称为胚状体的囊状结构，该结构由早期小鼠胚胎的典型组织谱系组成。因为胚状体维持血液发育，所以我们认为它们也可能支持原始生殖细胞的形成。在这里，我们从胚状体中分离出原始生殖细胞，并获得了不断增长的胚生殖细胞系。胚胎生殖细胞显示出Igf2r和H19基因的甲基化标记（烙印）的擦除，这是生殖谱系的特性。我们表明，胚状体支持原始生殖细胞成熟成单倍体雄配子，当注入卵母细胞时可恢复体细胞二倍体染色体互补并发育为胚泡。我们从胚胎干细胞衍生生殖细胞的能力为研究种系表观遗传修饰和哺乳动物配子发生提供了可访问的体外模型系统。

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