Identification of smooth muscle cells purified from rabbit bladder after in vitro culture / 复旦学报（医学版）

ABSTRACT

Objective To establish a method of obtaining the highly purified smooth muscle cells (SMCs) isolated from rabbit bladder and then to identify them after in vitro culture using methods of morphology and immunofluorescence. Methods Primary cell culture of SMCs was set up by enzymic method in total of 6 adult male New Zealand-White rabbits. The serosa and mucosa of bladder were carefully removed by micro-surgical instruments under the naked eyes. The morphology characteristics of cells and their pattern of cell growth were observed using inverted microscope. The cells were identified using methods of H-E staining, transmission electron microscope and immunofluorescence staining of α-actin. The curves of cell growth and proliferation were drawn after cell counting. Results The capability of adherence was observed in these isolated SMCs after in vitro culture for 24 hours, and passage was then conducted after culture for 7 days. The cells kept growing well and fast from the 1~(st) to 10~(th) generation. But starting from the 12~(th) generation, the deformed cells were found and then become those without typical morphological feature of SMCs at the 15~(th) generation, and finally died at the 20~(th) generation. The isolated cells after in vitro culture at 4~(th) passage were identified to be SMCs successfully using the methods of H-E staining, transmission electron microscope and immunofluorescence. Conclusions The highly purified bladder SMCs of rabbit were successfully obtained after isolated by enzymic method and in vitro culture within a short time. That would provide us the basis for future studies of pathophysiology and molecular biology of bladder SMCs.