Effective induction of inner ear hair cells from mouse embryonic stem cells

听力损失主要是由于耳蜗器或Corti器官中感觉毛细胞(HCs)的丧失造成的。虽然胚胎干细胞是一种很有前途的细胞治疗来源，但人们对如何从胚胎干细胞高效地产生类HC细胞知之甚少。在本研究中，我们建立了一种生长类胚体(EBS)的单一培养方法，其中使用ST2基质细胞的条件培养液(ST2-CM)培养14d的EBS。培养14天后，EB突起中高达20%的细胞表达与HC相关的标志，包括Math1(也称为atoh1)、myosin6、myosin7a、calretinin、α9AchR和Brn3c(也称为Pou4f3)，并显示出立体纤毛样结构的形成。此外，我们发现这些细胞在移植到鸡胚胎后被整合到发育中的内耳中。目前使用ST2-CM诱导内耳HC的方法(HIST2法)是一种简单、高效的方法，可以在较短的培养时间内获得ES来源的HC样细胞。

Hearing loss is mainly caused by loss of sensory hair cells (HCs) in the organ of Corti or cochlea. Although embryonic stem (ES) cells are a promising source for cell therapy, little is known about the efficient generation of HC-like cells from ES cells. In the present study, we developed a single-medium culture method for growing embryoid bodies (EBs), in which conditioned medium (CM) from cultures of ST2 stromal cells (ST2-CM) was used for 14-day cultures of 4-day EBs. At the end of the 14-day cultures, up to 20% of the cells in EB outgrowths expressed HC-related markers, including Math1 (also known as Atoh1), myosin6, myosin7a, calretinin, α9AchR and Brn3c (also known as Pou4f3), and also showed formation of stereocilia-like structures. Further, we found that these cells were incorporated into the developing inner ear after transplantation into chick embryos. The present inner ear HC induction method using ST2-CM (HIST2 method) is quite simple and highly efficient to obtain ES-derived HC-like cells with a relatively short cultivation time.