Subventricular zone microglia possess a unique capacity for massive in vitro expansion.
Subventricular zone microglia possess a unique capacity for massive in vitro expansion.
复制标题
室下区小胶质细胞具有独特的大规模体外扩张能力。
作者：
Marshall2nd,GregoryP;Demir,Meryem;Steindler,DennisA;Laywell,EricD
小胶质细胞是大脑的常驻免疫细胞，最近被假设在神经元疾病和年龄相关的神经性衰退中发挥作用，并且理论上是成人神经发生的调节剂。目前用于从培养的原代脑组织中分离小胶质细胞的方法导致相对较差的产率，需要大的组织样品或多个标本来获得足够数量的小胶质细胞用于细胞和分子分析。我们在这里报告了一种从已建立的胶质细胞单层培养物中重复分离小胶质细胞的方法，从中可以将小胶质细胞的初始群体扩增约10，000倍。扩增的群体表达适当的小胶质细胞形态和表型标志物，并显示功能正常的吞噬作用，从而为研究和分析来自初级组织的单一初始解剖的小胶质细胞提供了高产率测定。此外，这种大规模的扩增仅限于来自脑室下区的小胶质细胞，因为发现可分离的小胶质细胞的倍数扩增比来自其他脑区域的培养物高出20倍，这表明这种持续性神经源性区域的独特性质。© 2008 Wiley利斯公司
Microglia, the resident immune cells of the brain, have recently been hypothesized to play a role both in neuronal diseases and age‐related neurogenic decline, and are theorized to be modulators of adult neurogenesis. Current methods for the isolation of microglia from cultured primary brain tissue result in relatively poor yield, requiring a large tissue sample or multiple specimens to obtain a sufficient number of microglia for cell and molecular analysis. We report here a method for the repetitive isolation of microglia from established glial monolayer cultures from which it is possible to expand the initial population of microglia roughly 10,000‐fold. The expanded population expresses appropriate microglial morphology and phenotype markers, and demonstrates functionally normal phagocytosis, thus providing a high‐yield assay for the investigation and analysis of microglia from a single initial dissection of primary tissue. Furthermore, this massive expansion is limited to microglia derived from the subventricular zone as the fold expansion of isolatable microglia was found to be up to 20 times greater than cultures from other brain regions, indicating unique properties for this persistently neurogenic region. © 2008 Wiley‐Liss, Inc.