Establishment and characterization of rat progenitor hair cell lines
Establishment and characterization of rat progenitor hair cell lines
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DOI：
10.1016/s0378-5955(03)00077-7
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发表时间：
2003-05-01
期刊：
影响因子：
2.8
通讯作者：
Lin, JZ
中科院分区：
文献类型：
--
作者：
Ozeki, M;Duan, LJ;Lin, JZ
耳蜗祖细胞系是研究毛细胞发育过程中的细胞特性、基因表达特征和信号转导的有用细胞。为了获得胚胎和出生后耳蜗祖毛细胞系，我们将胚胎第12天(E12)的耳囊感觉神经上皮细胞原代培养和出生后第5天(P5)的Corti组织器官外植体永生化。将人乳头瘤病毒16型E6/E7基因导入原代培养细胞和外植体，传代50代，有限稀释法从上述P5器官分离出部分克隆性细胞。然后在这些细胞克隆中检测神经元、神经、上皮、毛细胞标记物和重要转录因子的表达。表达上述标记的克隆被认为是毛细胞的祖细胞。至少有两个具有代表性的细胞系，一个来自耳囊上皮细胞的混合培养，另一个来自Corti细胞器官，最终表达毛细胞标记和神经元/神经细胞标记。前者仅表达早期毛细胞标志物oncomodrin和myosin VIIa，而后者表达oncomodrin、calretinin、myosin VIIa和BRN3.1。这些细胞系可能代表了耳蜗发育不同阶段的前体毛细胞。(C)2003 Elsevier Science B.V.保留所有权利。
Cochlear progenitor hair cell lines are useful for studies of cellular specification, gene expression features, and signal transduction involved in the development of hair cells. To obtain embryonic and postnatal cochlear progenitor hair cell lines, we immortalized primary cultures of sensorineural epithelial cells from otocysts on embryonic day 12 (E12) and explants of the organ of Corti tissues on postnatal day 5 (P5). Primary cultures and explants were then transduced by the E6/E7 genes of human papilloma virus type 16. Transduced cells were passed for >50 passages and partial clonal cells were isolated from the above P5 organ of Corti explants by limiting dilution. The expression of neuronal, neural, epithelial, hair cell markers, and important transcription factors were then examined in these cell clones. Clones that express the above markers were considered as being progenitor hair cells. At least two representative cell lines, one from a mixed culture of otocyst epithelial cells and the other from the organ of Corti cells, ultimately expressed hair cell markers and neuronal/neural cell markers. The former only expressed the early hair cell marker oncomodulin and myosin VIIa, whereas the latter expressed oncomodulin, calretinin, myosin VIIa and Brn 3.1. These cell lines may represent progenitor hair cells at the different stages of cochlear development. (C) 2003 Elsevier Science B.V. All rights reserved.