大鼠主动脉内皮细胞改良型分离培养方法.

林翠红; 刘光辉; 杨田野; 赵 利; 王 航; 史常旋; 孟 春

Objective To establish a simple method of isolating, purifying and cultivating vascular endothelial cells (VECs) of rat aortas. Methods The aortas of rats were isolated and obtained under aseptic conditions. The intima of aortas were turned over and exposed using ophthalmic microscopic surgical instruments. The fragments of the aortas were divided into digesting-sheeting group (DSG), digesting group (DG), and sheeting group (SG) based on the different process modes, and then received cultivation. Morphological characterization was performed using phase contrast microscopy; further characterization was analyzed by immunofluorescence method. Results VECs migrated out of the fragments of aortas after 48-72 hours of plating, and the primary cells reached subconfluence on days 10-11. Their subcultures grew faster than the primary and reached confluence on day 5. The cells and their subpassages showed VECs' morphology with typical cobblestone appearance. The cells of DSG migrated from the fragments of aortas faster than those of DG and SG, and the quantity of primary cells of DSG were larger than those of DG and SG. Ninety-nine percent of the cell population had positive staining for factor VIII, von Willebrand factor and CD31. VECs cultured in this study could be passaged repeatedly for 10 passages without significant loss of typical features, and immunologically identified at the 8th generation without features changed. Conclusion We have developed a simple and economic method to isolate, purify and culture VECs of rat aortas.

Journal of Xi'an Jiaotong University (Medical Sciences), 2018, Vol 39, Issue 6, p911

1671-8259

Academic Journal

10.7652/jdyxb201806028