Isolation and culture of primary microvascular endothelial cells from mouse brains
Zhang Zuo-hui
Chen Chen
Chen Hao
Cui Gui-yun
Hua Fang
Abstract：BACKGROUND:Brain microvascular endothelial cel s (BMECs) are important tools in the field of neuroscience research;therefore, how to obtain highly purified BMECs is a key and difficulty in vitro. <br> OBJECTIVE:To develop a simple method of isolating and culturing highly purified BMECs. <br> METHODS:C57BL/6 mice aged 6-8 weeks old were selected, and microvessels were obtained using enzyme digestion and gradient centrifugation. Further, endothelia cel s were purified by certain drugs, fol owed by identified by CD31 and GFAP immunofluorescence staining. The expression of Claudin-5 was detected using immunofluorescence staining with anti-Claudin-5 antibody. <br> RESULTS AND CONCLUSION:Mouse BMECs grew and arranged in spiral or cobblestone-like. Immunofluorescence staining showed that the purity of BMECs reached above 99%and Claudin-5 was highly expressed. In conclusion, a simple method of easy accessibility is developed to obtain highly purified primary mouse BMECs.
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Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 7666-7671 )