Theus Michelle H, Ricard Jerome, Liebl Daniel J
The Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami, Florida, USA.
Curr Protoc Stem Cell Biol. 2012 Mar;Chapter 2:Unit 2D.8. doi: 10.1002/9780470151808.sc02d08s20.
Endogenous neural stem/progenitor cells (NSPCs) residing in the subventricular zone (SVZ) of the adult mouse forebrain have been shown to enhance their neurogenic potential in response to CNS injury. Mechanisms involved in regulating adult neurogenesis under naïve or stressed conditions can be studied using a monolayer cell-culture system of the nestin-expressing NSPC lineage to analyze proliferation, survival, and differentiation. Here, a protocol for the expansion of NSPCs for studies aimed at understanding the functional role of NSPCs in maintaining adult neurogenic processes is described. This unit outlines detailed procedures for: (1) isolation, maintenance, and culture of the NSPC component of the SVZ niche from the lateral wall of the lateral ventricle; (2) characterization of NSPC functions by examining proliferation, survival, and differentiation; and (3) efficient siRNA transfection methods in 96-well format.
成年小鼠前脑脑室下区（SVZ）中的内源性神经干细胞/祖细胞（NSPCs）已被证明在中枢神经系统损伤时会增强其神经发生潜能。在幼稚或应激条件下调节成体神经发生的机制，可以使用表达巢蛋白的NSPC谱系的单层细胞培养系统来研究，以分析增殖、存活和分化情况。本文描述了一种用于NSPCs扩增的方案，旨在了解NSPCs在维持成体神经发生过程中的功能作用。本单元概述了以下详细程序：（1）从侧脑室侧壁分离、维持和培养SVZ生态位的NSPC成分；（2）通过检查增殖、存活和分化来表征NSPC功能；（3）96孔板格式的高效siRNA转染方法。
