Deshpande Krutika, Saatian Behnaz, Martirosian Vahan, Lin Michelle, Julian Alex, Neman Josh
Department of Neurological Surgery, Keck School of Medicine, University of Southern California, Los Angeles, California.
Curr Protoc Stem Cell Biol. 2019 Jun;49(1):e80. doi: 10.1002/cpsc.80. Epub 2019 Feb 5.
A population of neural stem cells exists in the adult mammalian central nervous system. Purification and characterization of neurospheres provide valuable tools to study the regulation and differentiation of neural stem cells both in vitro and in vivo. Successful stimulation and production of neurospheres can ultimately be used for therapeutic purposes. The currently available methods are limited by their poor yield and the large number of animals required to compensate for that. Here, we describe a procedure to purify neurospheres from adult mouse whole brain. We provide detailed steps on how to propagate, passage, and maintain the adult neurospheres, and how to differentiate the pure neurospheres into the lineage of interest. Using this method, neurospheres can be easily derived from adult mouse whole brain. The derived adult neurospheres maintain their homogenous undifferentiated status while retaining their differentiation potential. This new protocol facilitates adult neurospheres isolation, purification, maintenance, and differentiation. © 2019 by John Wiley & Sons, Inc.
成年哺乳动物的中枢神经系统中存在一群神经干细胞。神经球的纯化和特性分析为研究神经干细胞在体外和体内的调控及分化提供了有价值的工具。成功刺激并产生神经球最终可用于治疗目的。目前可用的方法存在产量低以及需要大量动物来弥补这一缺陷的局限性。在此，我们描述了一种从成年小鼠全脑中纯化神经球的方法。我们提供了关于如何扩增、传代和维持成年神经球，以及如何将纯神经球分化为感兴趣谱系的详细步骤。使用这种方法，可以轻松地从成年小鼠全脑中获得神经球。所获得的成年神经球保持其均匀的未分化状态，同时保留其分化潜能。这个新方案有助于成年神经球的分离、纯化、维持和分化。© 2019 约翰威立父子出版公司
