[Culture and identification of rabbit cavernosal endothelial cells].
The objective of this study was to establish a protocol for isolating, culturing, proliferating cavernosal endothelial cells, and to obtain large number of autologous cavernosal endothelial cells for tissue-engineered substitutes. Cavernosal tissues were either minced into 1 to 2 mm3 pieces and incubated with 0.02% collagenase IV for 2h or first minced into 20 mm3 then incubated with 0.05% elastase for 1h, 2h or 3h, respectively; the undigested tissue blocks were repeatedly squeezed with a spatula for 2 min. Cells were collected and cultured with supplemented endothelial cell growth medium. Morphology and expansion of the cells were observed. Cell-type specific proteins CD31 and vWF were analyzed by immunohistochemical methods. The results revealed that cell isolation with 0.05% elastase for 2h followed by additional squeezing of residual tissue was effective not only for cell isolation, but also for preventing the tissue from getting contaminated by stromal cells, which was better than cell isolation with collagenase IV. Cavernosal endothelial cells were found with a typical cobblestone morphology and could be passaged 7 passages. Immunohistochemistry showed that endothelial specific proteins CD31 and vWF were positive. These results suggest that the isolation of cavernosal endothelial cells is simple and effective, the cells proliferate rapidly and they can be passaged easily. This method of new cultures will be of benefit to penile tissue engineering.
Wei
Yun
Y
Institute of Medical Virology, Medical College of Wuhan University, Wuhan 430071, China.
Wei
Lei
L
D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D008297:Male; D010413:Penis / Q000166:cytology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*; D014841:von Willebrand Factor / Q000378:metabolism
D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014841:von Willebrand Factor