中国药理学通报Issue(11)：1616-1619,4.DOI:10.3969/j.issn.1001-1978.2014.11.027
一种高纯度和高活力的大鼠脑微血管内皮细胞的提取及原代培养方法
A highly purified and active extract and primitive culture method for rat microvascular endothelial cells (BMECs)
摘要
Abstract
Aim To establish a highly purified,active and prac-tical extract and primitive culture method for rat brain microvas-cular endothelial cells ( BMECs) for providing materials for con-struction in vitro blood-brain barrier ( BBB) model. Methods Cerebral cortex of 1-2 week SD rats was collected,and successive digestion with typeⅡcollagenase and collagenase/dispase, sieve filtration and then twice gradient centrifugation in 20% BSA and 44% Percoll condition were used to obtain brain microvascular section. After that brain microvascular section was seeded in cul-ture bottle and then primarily cultured. Inverted microscope and factor-VIII relative antigen immunostaining methods were used for cellular morphological observation and identification. Results BMECs climbed out the vessel segment and proliferated with ad-herence after 2 h in vitro culture,and they became typical peb-bles structure after further 3-4 d culture. Morever, factor-VIII relative antigen immunostaining identified that expression for the endothelial cells was positive,cytoplasm was brown and positive cells account for more than 99%. Conclusion Rat BMECs with high purity could be extracted and cultured by using the above methods,and it has great potential for construction in vitro BBB model and in depth studies on biological characteristics and func-tions of BMECs.关键词
大鼠/脑微血管内皮细胞/原代培养/血脑屏障/形态学/免疫染色Key words
rat/brain microvascular endothelial cells/primitive culture/blood-brain barrier/morphology/Immunostaining分类
医药卫生引用本文复制引用
查雨锋,张顺,苏航,刘亭,傅晓钟,董永喜,王爱民,王永林..一种高纯度和高活力的大鼠脑微血管内皮细胞的提取及原代培养方法[J].中国药理学通报,2014,(11):1616-1619,4.基金项目
国家自然科学基金资助项目( No 81260473) （ No 81260473）
贵州省中药现代化科技产业研究开发专项项目( No 黔科合中药字2012-3013号) （ No 黔科合中药字2012-3013号）
贵州省优秀青年科技人才培养对象专项基金( No 2013-45号) （ No 2013-45号）
贵州省中药现代化科技产业研究开发专项(No 黔科合重G字[2013]4001) （No 黔科合重G字[2013]4001）
