Production of neurons, astrocytes and oligodendrocytes from mammalian CNS stem cells
Production of neurons, astrocytes and oligodendrocytes from mammalian CNS stem cells
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DOI：
10.1038/nprot.2008.55
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Weiss, Samuel
中科院分区：
文献类型：
--
作者：
Chojnacki, Andrew;Weiss, Samuel
在无血清培养系统中分离和扩增前体细胞可以系统地表征其特性以及调节其功能的内在和外在信号。成年小鼠大脑中神经干细胞的发现是通过创建一种新颖的培养系统（后来称为神经球测定）而成为可能的。其中，将分离的成年小鼠脑室周围区域在存在表皮生长因子但不存在粘附基质的情况下进行铺板，这导致增殖细胞球体的产生，这些细胞球体从板底部脱离并保持悬浮在培养基中。自问世以来，神经球培养系统已广泛应用于神经前体细胞领域，并广泛应用于角膜、心脏、皮肤、前列腺、乳腺和脑肿瘤干细胞的分离和扩增。原始神经球培养方案大约需要 10 天才能完成，此处详细描述。
The isolation and expansion of precursor cells in a serum-free culture system allows for the systematic characterization of their properties and the intrinsic and extrinsic signals that regulate their function. The discovery of neural stem cells in the adult mouse brain was made possible by the creation of a novel culture system subsequently termed the neurosphere assay. Therein, the dissociated adult mouse periventricular area was plated in the presence of epidermal growth factor, but in the absence of adhesive substrates, which resulted in the generation of spheres of proliferating cells that detached from the plate bottom and remained suspended in the media. Since its inception, the neurosphere culture system has been widely used in the neural precursor cell field and has been extensively adapted for the isolation and expansion of corneal, cardiac, skin, prostate, mammary and brain tumor stem cells. The original neurosphere culture protocol, which takes approximately 10d to complete, is described here in detail.