Isolation and culture of primary microvascular endothelial cells from mouse brains / 中国组织工程研究

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. OBJECTIVE:To develop a simple method of isolating and culturing highly purified BMECs. 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. 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.