Culture and identification of microglias and preoligodendrocytes
10.3760/cma.j.issn.1671-8925.2010.08.001
- VernacularTitle:小胶质细胞和少突胶质细胞前体的培养和鉴定
- Author:
Ya-Fang HE
1
;
Hui-Jin CHEN
;
Long-Hua QIAN
;
Guan-Yi CHEN
Author Information
1. 上海交通大学医学院附属新华医院
- Keywords:
Cell culture techniques;
Preoligodendrocyte;
Microglia
- From:
Chinese Journal of Neuromedicine
2010;09(8):757-760
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the isolated and in vitro cultural methods of preoligodendrocytes (preOLs) and microglias (MGs) obtained from the brain tissues of neonatal rats.Methods The MGs and preOLs isolated from the brain tissues of the 2-d-old SD neonatal rats were primarily cultured by using the nutrition-deficient method with the combination of shaking and the modified shaking and adherence method, respectively. The purity of the cultured cells was identified by immunocytochemical analysis. Results After cultured for 7 d, the mixed glias formed 3 cell layers consisting of the microglias in the upper layer, O2A progenitor cells in the middle layer and the astrocytes in the basal layer, respectively. It was observed under fluorescence microscope that the cultured preOLs were small and round with bi-polar or tri-polar prominence, and the cultured microglias displayed amebiform or round morphologies, sometimes with the burr rim shape. The immunocytochemical analysis identified that the purities of the cultures were consistently ＞95% for O4 positive labeled preOLs and ＞90% for FITC-labeled IB4 positive MG. Conclusion By using the nutrition-deficient method with the combination of shaking and the modified shaking and adherence method, the massive highly-pure and alive microglia and preOLs are obtained successfully.
