A Resource for the Transcriptional Signature of Bona Fide Trophoblast Stem Cells and Analysis of Their Embryonic Persistence
A Resource for the Transcriptional Signature of Bona Fide Trophoblast Stem Cells and Analysis of Their Embryonic Persistence
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DOI：
10.1155/2015/218518
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发表时间：
2015-01-01
影响因子：
4.3
通讯作者：
Arnold, Sebastian J.
中科院分区：
文献类型：
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作者：
Kuales, Georg;Weiss, Matthias;Arnold, Sebastian J.
滋养层干细胞（TSCs）代表产生胎盘胚胎部分不同细胞的多能祖细胞。在这里，我们分析了关键TSC转录因子Cdx 2，Eomes和Elf 5在小鼠胚胎早期发育胎盘和培养的TSC中的表达，并揭示了蛋白质水平的惊人异质性。我们分析了早期胎盘中TSCs的持续存在，发现TSCs保留在绒毛膜铰链中，直到E9.5，然后很快丢失。为了定义真正的TSC的转录特征，我们使用Eomes或Cdx 2和Eomes（GFP）的可诱导的功能获得和丧失等位基因来操纵和监测TSC的核心维持因子，然后进行全基因组表达谱分析。对所得表达谱的组合分析允许定义可能在功能上有助于维持TSC状态的新TSC标记基因。通过qRT-PCR和原位杂交的分析验证了新的TSC和绒毛膜特异性标记基因，如Bok/Mtd、Cldn 26、Duox 2、Duoxa 2、Nr 0 b1和Sox 21。因此，这些表达数据为真正的和早期分化的TSC的转录特征提供了有价值的资源，并且可以有助于增加对维持和/或建立TSC的干细胞性的转录电路的理解。
Trophoblast stem cells (TSCs) represent the multipotent progenitors that give rise to the different cells of the embryonic portion of the placenta. Here, we analysed the expression of key TSC transcription factors Cdx2, Eomes, and Elf5 in the early developing placenta of mouse embryos and in cultured TSCs and reveal surprising heterogeneity in protein levels. We analysed persistence of TSCs in the early placenta and find that TSCs remain in the chorionic hinge until E9.5 and are lost shortly afterwards. To define the transcriptional signature of bona fide TSCs, we used inducible gain- and loss-of-function alleles of Eomes or Cdx2, and Eomes(GFP), to manipulate and monitor the core maintenance factors of TSCs, followed by genome-wide expression profiling. Combinatorial analysis of resulting expression profiles allowed for defining novel TSC marker genes that might functionally contribute to the maintenance of the TSC state. Analyses by qRT-PCR and in situ hybridisation validated novel TSC- and chorion-specific marker genes, such as Bok/Mtd, Cldn26, Duox2, Duoxa2, Nr0b1, and Sox21. Thus, these expression data provide a valuable resource for the transcriptional signature of bona fide and early differentiating TSCs and may contribute to an increased understanding of the transcriptional circuitries that maintain and/or establish stemness of TSCs.