Abstract: BACKGROUND: To obtain a large number of high-purity endothelial cells, so as to carry out better experimental research on endothelial cell-related diseases, this paper updated the separation and culture methods of endothelial cells by improving the ratio of digestive enzymes.
OBJECTIVE: To explore the culture method of rabbit aortic endothelial cells in vitro and identify them, so as to lay a foundation for better experimental study of endothelial cell-related diseases.
METHODS: The endothelial tissue and cell suspension of rabbit aorta were obtained by compound enzyme digestion. After density gradient centrifugation, high-purity vascular endothelial cells were obtained and cultured in vitro. Furthermore, the rabbit aortic endothelial cells were identified by CD34 immunocytochemical staining and the cell purity was analyzed.
RESULTS AND CONCLUSION: (1) The vascular endothelial cells cultured in vitro were arranged in the shape of typical “paving stone”. (2) In addition, immunohistochemistry showed that the isolated cells were strongly positive for CD34-staining, to prove that they were endothelial cells. (3) The quantitative analysis of the immunohistochemical photos showed that all the cells were CD34-positive cells; that is, the purity of the obtained endothelial cells was more than 95%. (4) The endothelial tissue and cell suspension of rabbit aorta were obtained by compound enzyme digestion. After density gradient centrifugation, high-purity vascular endothelial cells were obtained. Rabbit aortic endothelial cells were isolated and cultured successfully in vitro. 

Key words: rabbit, aorta, endothelial cells, primary culture, identification, CD34, cell purity