Induction of inner ear hair cell-like cells from Math1-transfected mouse ES cells.
Induction of inner ear hair cell-like cells from Math1-transfected mouse ES cells.
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DOI：
10.1038/cddis.2013.230
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发表时间：
2013-07-04
影响因子：
9
通讯作者：
中科院分区：
文献类型：
--
作者：
Math 1是果蝇无调基因的一个基本螺旋-环-螺旋转录因子同源物，被认为是诱导Corti器或耳蜗发育过程中感觉毛细胞（HC）形成的关键因子。虽然胚胎干细胞（ES）能够产生HC样细胞，但Math 1在诱导这些细胞中的作用尚未完全阐明。在本研究中，我们将Math 1导入ES细胞，以实现HC样细胞的高效产生。使用Tet-On基因表达系统产生携带Tet-inducible Math 1的ES细胞，Math 1-ES细胞。使在不存在多西环素（Dox）4天的情况下形成的胚状体（EB）在400 μg/ml Dox的存在下在培养皿中再生长14天。在这些14天的培养结束时，EB生长物中大约10%的细胞表达HC相关标记物肌球蛋白6、肌球蛋白7a、钙视网膜蛋白、α 9 AchR和Brn 3c（也称为Pou 4f 3），并显示出静纤毛样结构的形成，而在没有Dox的EB生长物中很少有细胞显示出这些标记物。使用Brn 3c启动子质粒的Math 1-ES细胞的报告基因测定证明Math 1对Brn 3c具有正调控作用。此外，发现来自Math 1-ES细胞的这种HC相关标记阳性细胞在移植到鸡胚中后被掺入发育中的内耳中。Math 1-ES细胞被认为是ES衍生的HC样细胞的有效来源，Math 1可能是从分化的ES细胞诱导HC样细胞的重要因素。
Math1, a basic helix-loop-helix transcription factor homolog of the Drosophila atonal gene, is considered to be a key factor for induction of sensory hair cells (HCs) during development of the organ of Corti or cochlea. Although embryonic stem (ES) cells are able to produce HC-like cells, the role of Math1 in induction of those cells has not been thoroughly elucidated. In the present study, we introduced Math1 into ES cells in order to achieve efficient generation of HC-like cells. ES cells carrying Tet-inducible Math1, Math1-ES cells, were generated using a Tet-On gene expression system. Embryoid bodies (EBs) formed in the absence of doxycycline (Dox) for 4 days were allowed to grow for an additional 14 days in the dishes in the presence of 400 μg/ml of Dox. At the end of those 14-day cultures, approximately 10% of the cells in EB outgrowths expressed the HC-related markers myosin6, myosin7a, calretinin, α9AchR, and Brn3c (also known as Pou4f3) and showed formation of stereocilia-like structures, whereas few cells in EB outgrowths grown without Dox showed those markers. Reporter assays of Math1-ES cells using a Brn3c-promoter plasmid demonstrated positive regulation of Brn3c by Math1. Furthermore, such HC-related marker-positive cells derived from Math1-ES cells were found to be incorporated in the developing inner ear after transplantation into chick embryos. Math1-ES cells are considered to be an efficient source of ES-derived HC-like cells, and Math1 may be an important factor for induction of HC-like cells from differentiating ES cells.
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影响因子：
2.8
作者：
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通讯作者：
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