Epithelial-mesenchymal transition in colonies of rhesus monkey embryonic stem cells: A model for processes involved in gastrulation
Epithelial-mesenchymal transition in colonies of rhesus monkey embryonic stem cells: A model for processes involved in gastrulation
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DOI：
10.1634/stemcells.2004-0234
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发表时间：
2005-06-01
期刊：
影响因子：
5.2
通讯作者：
Thie, M
中科院分区：
文献类型：
--
作者：
Behr, R;Heneweer, C;Thie, M
恒河猴胚胎干细胞（rhES）在小鼠胚胎成纤维细胞（MEF）饲养层上生长长达10天，形成多层集落。在此期间，干细胞集落短暂分化成具有盘状形态的复杂结构。通过形态学、免疫组织化学和标记物mRNA表达对这些复杂集落进行表征，以鉴定上皮形成以及上皮-间充质转化（EMT）和模式形成的过程。典型地，分化的集落由上和下ES细胞层组成，前者生长在MEF细胞层的顶部，而下ES细胞层在MEF细胞下面展开。有趣的是，在殖民地的中部，有一个圆形的坑。开发在这里，饲养层消失，上层细胞似乎进入并通过凹陷向下迁移以形成下层，同时经历从上皮向间充质表型的转变，这由下层中标志蛋白E-钙粘蛋白和ZO-1的丢失指示。为了支持这一点，我们发现了一个伴随的10倍上调的基因蜗牛2，这是一个关键的调节EMT过程。中胚层标记Brachyury的调节表达也表明了上胚层向中胚层的转化。EMT是脊椎动物原肠胚形成的特征过程。因此，这些rhES细胞集落可能是一个有趣的模型，研究在灵长类动物早期胚胎发生的一些基本过程，并可能开辟新的途径，在体外研究EMT的调控。
Rhesus monkey embryonic stem (rhES) cells were grown on mouse embryonic fibroblast (MEF) feeder layers for up to 10 days to form multilayered colonies. Within this period, stern cell colonies differentiated transiently into complex structures with a disc-like morphology. These complex colonies were characterized by morphology, immunohistochemistry, and marker mRNA expression to identify processes of epithelialization as well as epithelial-mesenchymal transition (EMT) and pattern formation. Typically, differentiated colonies were comprised of an upper and a lower ES cell layer, the former growing on top of the layer of MEF cells whereas the lower ES cell layer spread out underneath the MEF cells. Interestingly, in the central part of the colonies, a roundish pit. developed. Here the feeder layer disappeared, and upper layer cells seemed to ingress and migrate through the pit downward to form the lower layer while undergoing a transition from the epithelial to the mesenchymal phenotype, which was indicated by the loss of the marker proteins E-cadherin and ZO-1 in the lower layer. In support of this, we found a concomitant 10-fold upregulation of the gene Snail2, which is a key regulator of the EMT process. Conversion of epiblast to mesoderm was also indicated by the regulated expression of the mesoderm marker Brachyury. An EMT is a characteristic process of vertebrate gastrulation. Thus, these rhES cell colonies may be an interesting model for studies on some basic processes involved in early primate embryogenesis and may open new ways to study the regulation of EMT in vitro.