Find in Library

Isolation method to obtain pure BMVECs is hard to be done consistently and remains a challenge. In this study, we isolated BMVECs from Wistar rat and C57/Bl6 mouse from Japan SLC. All procedures performed according to guidelines for the care and use of laboratory animals of Gunma University, Japan. The modification of isolation method was using bovine serum albumin (BSA) gradation, not Dextran-70 in which generally used, to separate clusters of BMVECs into single cell. This study was done at Universitas Padjadjaran, in colaboration with Gunma University, Japan, January 2008–June 2009. Further,characteristic and purification results were proven by imunofluorescene staining. The results showed that staining of tight junction, ZO-1, formed a monolayer, tightly packed, non-overlapping and contact-inhibited BMVECs, as expected for a vessel wall endothelial. ECs phenotype confirmed by acethylated LDL, von Willebrand and CD31. The digestion of capillaries generated contaminating pericytes. Contamination was purified using puromycin and the results considered satisfactory (98.3%). In conclusion, our modification procedure allows the isolation of primary rat and mouse BMVECs, which form an endothelial-like monolayer in few days. Puromycin can be used for purification of primary rat and mouse BMVECs