Isolation of sphere-forming stem cells from the mouse inner ear.
Isolation of sphere-forming stem cells from the mouse inner ear.
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DOI：
10.1007/978-1-59745-523-7_9
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Heller, Stefan
中科院分区：
文献类型：
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作者：
Oshima, Kazuo;Senn, Pascal;Heller, Stefan
哺乳动物内耳再生失去的感觉毛细胞的能力非常有限。当毛细胞损失导致听力损失时，这种缺陷变得明显，这是耳毒性损伤或衰老过程的结果。巧合的是，由于失去了再生毛细胞的能力，成年耳蜗似乎没有具有增殖能力的细胞，这些细胞可以作为失去细胞的祖细胞。相比之下，成年哺乳动物前庭感觉上皮显示有限的毛细胞再生能力，并且可以从成年小鼠前庭系统分离具有干细胞特征的球形成细胞。然而，新生儿内耳确实存在于耳蜗和前庭组织中的球形干细胞。在这里，我们提供的协议，以分离球形成干细胞从新生儿前庭和耳蜗感觉上皮细胞，以及从螺旋神经节。我们进一步描述了球体繁殖，细胞分化和表征的内耳细胞类型来自球体的程序。来自小鼠内耳的球体形成干细胞是受损内耳细胞替代策略发展的重要工具，并且是移植研究的真正祖细胞来源。
The mammalian inner ear has very limited ability to regenerate lost sensory hair cells. This deficiency becomes apparent when hair cell loss leads to hearing loss as a result of either ototoxic insult or the aging process. Coincidently, with this inability to regenerate lost hair cells, the adult cochlea does not appear to harbor cells with a proliferative capacity that could serve as progenitor cells for lost cells. In contrast, adult mammalian vestibular sensory epithelia display a limited ability for hair cell regeneration, and sphere-forming cells with stem cell features can be isolated from the adult murine vestibular system. The neonatal inner ear, however, does harbor sphere-forming stem cells residing in cochlear and vestibular tissues. Here, we provide protocols to isolate sphere-forming stem cells from neonatal vestibular and cochlear sensory epithelia as well as from the spiral ganglion. We further describe procedures for sphere propagation, cell differentiation, and characterization of inner ear cell types derived from spheres. Sphere-forming stem cells from the mouse inner ear are an important tool for the development of cellular replacement strategies of damaged inner ears and are a bona fide progenitor cell source for transplantation studies.