大鼠阴茎海绵体内皮细胞的分离培养与鉴定.

Objective Study to establish an in vitro method for the isolation, culture and identification of endothelial cells from the penile corpus cavernosum of SD rats. Methods The endothelial cells were purified by collagenase digestion and mechanical curettage. After 11 days of culture in EGM-2 complete culture medium, the cells were subcultured for the second generation. After hematoxylin-eosin staining, the endothelial cells were observed under light microscope, and the cell growth curve was drawn by CCK8 method. The expression of CD31 and vWF in endothelial cells was identified by immunofluorescence method, the cycle changes of endothelial cells were detected by flow cytometry, and the expression changes of vWF marker in endothelial cells were observed by laser confocal microscopy. Results The endothelial cells of rat penile corpus cavernosum isolated by collagenase digestion covered more than 2/3 of the culture flask area in 11 ~14d. After HE staining, the cells showed typical paving stone shape under light microscope. The co-positive expression rate of CD31 and vWF was (86. 19±1. 14)%. VWF could be stably expressed in the first 4 generations of cells and the growth cycle of the second and third generations of cells was stable. After hematoxylin-eosin staining, the cells appear typical pavement-like under light microscopy. Conclusion High purity endothelial cells from rat penile corpus cavernosum can be successfully isolated by collagenase digestion combined with mechanical scraping,which can be used as a cellular model for the study of endothelial cells in the penile corpus cavernosum. [ABSTRACT FROM AUTHOR]