Separation, culture and identification of SD rat corpus cavernosal endothelial cells

Abstract

The aim of the study is to investigate the methods of separation, culture and identification of Sprague Dawley (SD) rat corpus cavernosal vascular endothelial cells (CCECs). Cavernosal tissues were isolated from male SD rats. Enzymatic digestion was applied to separate CCECs. Purified cells were obtained using immunomagnetic beads and flow cytometric cell sorting and subcultured in EMG-2 medium. The growth curve of CCECs was measured by the tetrazolium salt 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The cells were identified by von Willebrand factor (vWF) using immunofluorescence, and the positive percentage of vWF expression was detected by flow cytometry. The monomorphic cobblestone-like cells were observed by microscopy. High purification was obtained using immunomagnetic beads. After 2 days of incubation, cells entered the logarithmic growth phase and reached a plateau on the fifth day. The vWF expression in cytoplasm was positive. The purity of cells was 95.8%, which was tested by flow cytometry. SD rat CCECs can be separated and cultured successfully by the method of enzymatic digestion, immunomagnetic beads and flow cytometric cell sorting.