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Endoderm Complexity in the Mouse Gastrula Is Revealed Through the Expression of Spink3

机译：通过Spink3的表达揭示小鼠内胚层的内胚层复杂性。

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Endoderm formation in the mammalian embryo occurs first in the blastocyst, when the primitive endoderm and pluripotent cells resolve into separate lineages, and again during gastrulation, when the definitive endoderm progenitor population emerges from the primitive streak. The formation of the definitive endoderm can be modeled using pluripotent cell differentiation in culture. The differentiation of early primitive ectoderm-like (EPL) cells, a pluripotent cell population formed from embryonic stem (ES) cells, was used to identify and characterize definitive endoderm formation. Expression of serine peptidase inhibitor, Kazal type 3 ( Spink3 ) was detected in EPL cell–derived endoderm, and in a band of endoderm immediately distal to the embryonic–extra-embryonic boundary in pregastrula and gastrulating embryos. Later expression marked a region of endoderm separating the yolk sac from the developing gut. In the embryo, Spink3 expression marked a region of endoderm comprising the distal visceral endoderm, as determined by an endocytosis assay, and the proximal region of the definitive endoderm. This region was distinct from the more distal definitive endoderm population, marked by thyrotropin-releasing hormone ( Trh ). Endoderm expressing either Spink3 or Trh could be formed during EPL cell differentiation, and the prevalence of these populations could be influenced by culture medium and growth factor addition. Moreover, further differentiation suggested that the potential of these populations differed. These approaches have revealed an unexpected complexity in the definitive endoderm lineage, a complexity that will need to be accommodated in differentiation protocols to ensure the formation of the appropriate definitive endoderm progenitor in the future.
机译：哺乳动物胚胎中的内胚层形成首先发生在胚泡中，这时原始的内胚层和多能细胞分解成不同的谱系，然后在成胃过程中再次出现，当确定的内胚层祖细胞从原始条纹中出现时。可以使用培养中的多能细胞分化来模拟定形内胚层的形成。早期原始外胚层样（EPL）细胞的分化是由胚胎干（ES）细胞形成的多能细胞群，用于鉴定和表征定形内胚层的形成。在EPL细胞衍生的内胚层中，以及在前胚和胚化胚胎的胚-胚外边界附近的一段内胚层中检测到了丝氨酸肽酶抑制剂Kazal 3型（Spink3）的表达。后来的表达标志着将卵黄囊与发育中的肠分开的内胚层区域。在胚胎中，Spink3表达标记了一个内胚层区域，该区域包括通过内吞作用测定确定的远端内脏内胚层和定形内胚层的近端区域。该区域与更远端的定形内胚层种群不同，其特征在于促甲状腺激素释放激素（Trh）。在EPL细胞分化过程中可能形成表达Spink3或Trh的内胚层，而这些种群的患病率可能受到培养基和生长因子添加的影响。此外，进一步的分化表明这些人群的潜力不同。这些方法揭示了定形内胚层谱系中出乎意料的复杂性，为确保将来形成合适的定形内胚层祖细胞，必须将这些复杂性纳入分化方案中。

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