BACKGROUND:Primary vascular endothelial cels are mostly harvested through aorta endothelial cel cultures and micro-artery endothelial cel cultures using enzyme digestion and tissue adhesion methods, and the quality and purity of harvested cels cannot meet the need for current scientific research.
OBJECTIVE:To investigate an improved extraction of primary vascular endothelial cels and the relevant identification method.
METHODS: A segment of rabbit aorta was cut to culture vascular endothelial cels using the improved extraction method in group A or using adhesion method in group B. In the group A, the vascular intima was striped out with microsurgical instruments, and digested enzymaticaly to acquire single primary cels folowed by culture in endothelial cel culture medium. In the group B, the whole vascular intima was adhered to the culture dish that was incubated in a 5%CO2, 37℃ incubator for 1 hour. Cel pelets in the two groups were culturedin vitro. Cel morphology was observed using a microscope; immunohistochemical staining was used to detect CD31, VIII factor and Vimentin protein for identification of vascular endothelial cels.
RESULTS AND CONCLUSION:The purity and number of vascular endothelial cels extracted by the improved method were higher than those by the adhesion method. Immunohistochemical findings showed positive expression of CD31 and VIII factor, but negative expression of Vimentin. These findings indicate that the improved extraction method can obtain more vascular endothelial cels with higher purity, which is of strong operability and practicality.