In Vitro Differentiation of Inner Ear Hair Cells

描述(申请人提供)：成人感音神经性耳聋 主要是由于内耳感觉毛细胞的永久性丧失。在……里面 在这方面，哺乳动物不同于低等脊椎动物，后者可以再生感觉 损伤后邻近支持细胞的毛细胞。目前还不清楚是否 这种差异是由于儿童缺乏感觉发育能力。 哺乳动物的支持细胞，或由于哺乳动物缺乏某些内源性刺激 刺激耳部损伤后的再生。本文件中描述的实验 一项提案将标记胚胎的感觉并支持细胞前体 荧光转基因。这些前体将从胚胎中提纯出来 荧光激活细胞分选(FACS)和体外分化，使用一种 最近发展起来的培养体系。该系统还将用于隔离和 胚胎和出生后有丝分裂后支持细胞的分化 动物。从这些实验中，将确定有丝分裂后 哺乳动物的支持细胞能够产生感觉毛细胞，如果是的话，是如何产生的 在长期的发展过程中，这种能力依然存在。

DESCRIPTION (provided by applicant): Sensorineural deafness in adults is primarily due to the permanent loss of sensory hair cells in the inner ear. In this regard, mammals differ from lower vertebrates, which can regenerate sensory hair cells from adjacent supporting cells after injury. It is not known whether this difference arises from a lack of sensory developmental capacity in mammalian support cells, or from a lack of some endogenous stimulus in mammals to provoke regeneration after otic injury. Experiments described in this proposal will mark embryonic sensory and support cell precursors with a fluorescent transgene. These precursors will be purified from embryos using fluorescence activated cell sorting (FACS) and differentiated in vitro, using a recently developed culture system. This system will also be used to isolate and differentiate post-mitotic support cells from embryos and from post-natal animals. From these experiments, it will be determined whether post-mitotic mammalian support cells are able to generate sensory hair cells, and if so, how long in development this capacity persists.