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目的探讨兔阴茎海绵体平滑肌细胞快速分离方法,为应用膜片钳技术研究阴茎勃起机制提供实验材料。方法采用木瓜蛋白酶和胶原酶两步酶解消化法,快速分离出新西兰大白兔阴茎海绵体平滑肌细胞并应用免疫组化鉴定。结果分离的细胞成活率较高,贴壁呈长梭形,胞膜光滑完整,胞浆均匀,可用于膜片钳记录。免疫组化鉴定为兔阴茎海绵体平滑肌细胞。结论酶解消化快速分离兔阴茎海绵体平滑肌细胞为全细胞膜片钳技术研究阴茎勃起功能障碍的电生理机制提供了较好的实验材料。
Objective To investigate the rapid separation method of corpus cavernosum smooth muscle cells in rabbits and to provide experimental materials for the study of penile erection mechanism using patch clamp technique. Methods papain and collagenase two-step enzymatic digestion method, rapid isolation New Zealand white rabbit corpus cavernosum smooth muscle cells and identified by immunohistochemistry. Results The isolated cells had a higher survival rate, a long fusiform adherent wall, a smooth and intact cell membrane, and a uniform cytoplasm that could be used for patch-clamp recording. Immunohistochemistry was identified as rabbit corpus cavernosum smooth muscle cells. Conclusion Enzymatic digestion and rapid isolation of rabbit corpus cavernosum smooth muscle cells for whole-cell patch clamp technique to study the electrophysiological mechanism of penile erection provides a good experimental material.