ISOLATION AND FLOW CYTOMETRIC CHARACTERIZATION OF NEWBORN MOUSE BRAIN-DERIVED MICROGLIA MAINTAINED INVITRO
ISOLATION AND FLOW CYTOMETRIC CHARACTERIZATION OF NEWBORN MOUSE BRAIN-DERIVED MICROGLIA MAINTAINED INVITRO
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DOI：
10.1002/jlb.50.1.86
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发表时间：
1991-07-01
影响因子：
5.5
通讯作者：
DOUGLAS, SD
中科院分区：
文献类型：
--
作者：
HASSAN, NF;RIFAT, S;DOUGLAS, SD
用不同的小鼠巨噬细胞标记物在发育中和成年小鼠的脑白质中发现了小胶质细胞。虽然已经描述了几种分离小鼠小胶质细胞的技术，但小细胞产量和部分纯化限制了这些研究的进展。我们现在描述使用改进的McCarthy和De Vellis方法分离小鼠小胶质细胞。1~2日龄新生小鼠脑组织机械分离和化学分离，体外培养保存3周。在原代分离的脑培养中，观察到小胶质细胞从小集落迁移10天后出现。培养16~20天后，连续摇床培养16小时，可获得高细胞产率的脑源性小胶质细胞。与常驻的小鼠腹膜巨噬细胞相比，流式细胞术显示小胶质细胞表达较少的II类(Ia)抗原和少量的L3T4(CD4)抗原。
Microglia have been identified in the white matter of developing and adult mouse brain using different murine macrophage markers. While several techniques for the isolation of murine microglia have been described, the small cell yields and partial purification have limited the progress of these studies. We now describe the isolation of murine microglia using a modification of McCarthy and de Vellis method. Brain tissues from 1-2 day old newborn mice were mechanically and chemically dissociated and maintained in in vitro culture for 3 weeks. In primary dissociated brain cultures, microglia are observed after 10 days migrating from small colonies. After 16-20 days, brain-derived microglia were isolated with high cell yields by continuous shaking of the cultures for 16 hr. In contrast to resident murine peritoneal macrophages, microglia express less Class II (Ia) antigen and a small percentage express L3T4 (CD4) antigen by flow cytometry.