Isolation and Cultivation of Primary Brain Endothelial Cells from Adult Mice

Bio Protoc. 2017 May 20;7(10):e2294. doi: 10.21769/BioProtoc.2294.


Brain endothelial cells are the major building block of the blood-brain barrier. To study the role of brain endothelial cells in vitro, the isolation of primary cells is of critical value. Here, we describe a protocol in which vessel fragments are isolated from adult mice. After density centrifugation and mild digestion of the fragments, outgrowing endothelial cells are selected by puromycin treatment and grown to confluence within one week.

Keywords: Blood-brain barrier; CD31; Claudin-5; Occludin; Primary culture; Tight junctions; VE-cadherin; ZO-1.