Methods of Culturing and Identifying Brain Microvascular Endothelial Cells in Rats

WU Xiu-qin; MEI Xiao-yun; WU Hao-xin; LI Yang; LI Hui

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Methods of Culturing and Identifying Brain Microvascular Endothelial Cells in Rats

更新时间：2024-01-04

- Methods of Culturing and Identifying Brain Microvascular Endothelial Cells in Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 17, Issue 3, Pages: 187-189(2011)
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- Published：2011
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WU Xiu-qin, MEI Xiao-yun, WU Hao-xin, et al. Methods of Culturing and Identifying Brain Microvascular Endothelial Cells in Rats[J]. Chinese journal of experimental traditional medical formulae, 2011, 17(3): 187-189.
DOI：

WU Xiu-qin, MEI Xiao-yun, WU Hao-xin, et al. Methods of Culturing and Identifying Brain Microvascular Endothelial Cells in Rats[J]. Chinese journal of experimental traditional medical formulae, 2011, 17(3): 187-189. DOI：

摘要

目的: 探索纯度较高、相对经济简便的大鼠脑微血管内皮细胞分离和原代培养的方法。方法: 取1~10 d SD大鼠脑皮质经胰蛋白酶消化后

通过不同孔径的筛网过滤

再用胶原酶2次消化获得的微血管内皮细胞进行培养

采用相差显微镜形态学观察及Ⅷ因子相关抗原免疫组化鉴定。结果: 1~2 d细胞呈区域性单层贴壁生长

7~9 d呈典型的铺路卵石样征象

Ⅷ因子相关抗原免疫组化检测内皮细胞表达阳性

可见细胞的胞浆及核周呈棕黄色

胞核呈阴性

阳性细胞占绝大部分。结论: 采用2次消化、2次过滤技术可以成功分离培养出较理想的大鼠脑微血管内皮细胞。

Abstract

Objective: To explore the methods of culturing highly purified rat brain microvascular endothelial cells(BMEC). Method: BMEC of SD rats were prepared by filtering through a mesh

and digested by collagenase. Then the cells were examined under microscope and the expression of factor Ⅷ polyclone antibody was determined. Result: The cells became polygon arranging as single layer in 1-2 days. In 7-10 days

the cells arranged as ovum-like in compact monolayer. Immunocytochemistry of factor Ⅷ polyclone antibody showed positive in cytomembrane and cytoplasm

but negative in cell nucleus. Most of the cells were found to express factor Ⅷ-related antigen. Conclusion: Ideal rat brain microvascular endothelial cells can be cultured by the improved cultural method of double filtering and digestion.

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