Culture and purification of SD rat corpus cavernosum endothelial cells by enzymatic digestion combined with mechanical extrusion and fixed‐point digestion

• Published in: Andrologia Vol. 53; no. 10; pp. e14194 - n/a
• Main Authors: Chen, Ying, Qi, Tao, Zhu, Shu‐Guang, Li, Hao, Feng, Jia‐Xin, Zhang, Bin, Li, Shi‐Xiong, Ma, Shuai, Ma, Qiang, Chu, Qing‐Jun, Yang, Wen‐tao, Chen, Jun
• Format: Journal Article
• Language: English
• Published: Berlin Wiley Subscription Services, Inc 01.11.2021
• Subjects: cavernous endothelial cell; Cell culture; Collagen; Collagenase; Elastase; Endothelial cells; enzymatic digestion; Erectile dysfunction; fixed‐point digestion; Flow cytometry; Immunofluorescence; mechanical extrusion; Penis; Purification
• ISSN: 0303-4569; 1439-0272
• DOI: 10.1111/and.14194

To explore a new method of in vitro culture and purification of rat corpus cavernosum endothelial cells (CCECs). Male Sprague‐Dawley rats' penile tissue were digested with elastase or collagenase combined with mechanical extrusion to isolate and culture the CCECs. The fixed‐point digestion method was used to purify the primary cells. High‐purity CCECs were successfully isolated. Following the digestion of the primary CCECs by elastase or collagenase coupled with mechanical extrusion, the cells were paving stone‐ and cobblestone‐shaped over 10 days. The cell purity yielded in the second generation (P2) CCECs after using the fixed‐point digestion method was significantly high. Compared with primary CCECs extracted by elastase digestion combined with the mechanical extrusion method, CCECs cultured by collagenase digestion yielded higher purity and a more stable morphology after fixed‐point digestion and purification. Immunofluorescence staining of the third generation CCECs and the expression results of endothelial cell‐associated marker antibodies CD31 and VWF were positive, and flow cytometry showed the purity of CCECs was 96.9%. Enzymatic digestion combined with mechanical extrusion and fixed‐point digestion is a simple, economical method for in vitro culture and purification of CCECs, which is conducive to studying the pathophysiological mechanisms of endothelial dysfunction and erectile dysfunction.