Galli Rossella
Neural Stem Cell Biology Unit, Division of Neuroscience, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Methods Mol Biol. 2025;2905:233-244. doi: 10.1007/978-1-0716-4418-8_15.
The discovery of neural stem cells (NSCs) in the mammalian brain has raised many expectations as these unique cells might recapitulate different neurological diseases, including brain tumors, both from a functional and molecular perspective. Proper in vitro culturing of NSCs has emerged as a critical methodological issue, given that it should preserve the in vivo features of NSCs, with particular emphasis on cell heterogeneity. At the same time, the methodology for NSC culturing should allow the production of large amounts of cells to be exploited not only for prospective clinical applications, but also for drug screening. Direct in vitro selection of NSCs and, very recently, cancer stem cells (CSCs) by means of defined serum-free conditions represents the most reliable methodology to obtain long-term expanding SC lines. Here we describe the methods currently employed to enrich for NSCs/CSCs based on the NeuroSphere Assay (NSA) and their adaptation to specific assays for testing the efficacy of neuroactive compounds.
哺乳动物脑中神经干细胞（NSCs）的发现引发了诸多期待，因为这些独特的细胞可能从功能和分子角度重现包括脑肿瘤在内的不同神经疾病。鉴于NSCs的体外培养应保留其体内特征，尤其是细胞异质性，因此NSCs的适当体外培养已成为一个关键的方法学问题。与此同时，NSC培养方法应能够产生大量细胞，不仅用于预期的临床应用，还用于药物筛选。通过特定的无血清条件直接在体外选择NSCs，以及最近选择癌症干细胞（CSCs），是获得长期扩增干细胞系的最可靠方法。在此，我们描述了目前基于神经球测定法（NSA）富集NSCs/CSCs所采用的方法，以及它们如何适用于测试神经活性化合物功效的特定测定。
