Multipotent cell fate of neural crest-like cells derived from embryonic stem cells
Multipotent cell fate of neural crest-like cells derived from embryonic stem cells
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DOI：
10.1634/stemcells.2006-0323
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发表时间：
2007-02-01
期刊：
影响因子：
5.2
通讯作者：
Kunisada, Takahiro
中科院分区：
文献类型：
--
作者：
Motohashi, Tsutomu;Aoki, Hitomi;Kunisada, Takahiro
神经嵴细胞在整个胚胎中迁移并分化成不同的衍生物：外周神经元、颅间充质细胞和黑素细胞。由于神经嵴细胞在器官发生中具有重要作用，因此详细阐明神经嵴细胞的分化在发育生物学中具有重要意义。我们最近报道了黑素细胞可以从小鼠胚胎干细胞诱导。在这里，我们改进了培养系统，并显示了神经嵴样前体的存在。在培养液中添加视黄酸可降低造血功能并促进神经嵴标记基因的表达。形成的集落含有神经嵴细胞衍生物：神经元和神经胶质细胞，以及黑素细胞。这表明，神经嵴样细胞假设多个细胞的命运已经产生在这些目前的文化。为了分离神经嵴样细胞，我们分析了c-Kit的表达，c-Kit是一种在体内神经嵴细胞早期表达的细胞表面蛋白。c-Kit阳性（c-Kit（+））细胞早在培养期第9天就出现，并表达在神经嵴细胞中表达的转录因子Sox 10和Snail。当c-Kit（+）细胞从培养物中分离出来并重新培养时，它们经常形成含有神经元、神经胶质细胞和黑素细胞的集落。甚至单个c-Kit（+）细胞也形成了包含这三种细胞类型的集落，证实了它们的多潜能细胞命运。c-Kit（+）细胞也能够在体内沿沿着神经嵴迁移途径迁移。这些结果表明，c-Kit（+）细胞分离自黑素细胞分化培养的胚胎干细胞是密切相关的神经嵴细胞。
Neural crest cells migrate throughout the embryo and differentiate into diverse derivatives: the peripheral neurons, cranial mesenchymal cells, and melanocytes. Because the neural crest cells have critical roles in organogenesis, detailed elucidation of neural crest cell differentiation is important in developmental biology. We recently reported that melanocytes could be induced from mouse ESCs. Here, we improved the culture system and showed the existence of neural crest-like precursors. The addition of retinoic acid to the culture medium reduced the hematopoiesis and promoted the expression of the neural crest marker genes. The colonies formed contained neural crest cell derivatives: neurons and glial cells, together with melanocytes. This suggested that neural crest-like cells assuming multiple cell fates had been generated in these present cultures. To isolate the neural crest-like cells, we analyzed the expression of c-Kit, a cell-surface protein expressed in the early stage of neural crest cells in vivo. The c-Kit-positive (c-Kit(+)) cells appeared as early as day 9 of the culture period and expressed the transcriptional factors Sox10 and Snail, which are expressed in neural crest cells. When the c-Kit(+) cells were separated from the cultures and recultured, they frequently formed colonies containing neurons, glial cells, and melanocytes. Even a single c-Kit(+) cell formed colonies that contained these three cell types, confirming their multipotential cell fate. The c-Kit(+) cells were also capable of migrating along neural crest migratory pathways in vivo. These results indicate that the c-Kit(+) cells isolated from melanocyte-differentiating cultures of ESCs are closely related to neural crest cells.