兔膀胱平滑肌细胞的分离、培养和鉴定

A enzymic method for the isolation and culture of rabbit bladder smooth muscle cells

目的:建立兔膀胱平滑肌细胞的分离、培养和鉴定的方法。 方法:成年新西兰白兔2只(正常及梗阻各1 只),采用酶法分离技术获得膀胱平滑肌细胞后,于10%小牛血清的DMEM中培养,观察细胞形态和扩增情况,用 爬片染色、电镜、蛋白质α 肌动蛋白(α actin)鉴定细胞类型。 结果:在倒置显微镜下观察均呈“谷和峰”样结构、 细胞爬片苏木精 伊红(H E)染色及电镜检查均证实为平滑肌细胞。免疫组化染色检测α actin呈阳性反应。结果 证明,用该方法所分离、培养的膀胱平滑肌细胞其纯度几乎达99%。 结论:该方法易行、可靠,在短期内可获得大 量高纯度膀胱平滑肌细胞。

Objective:To establish a method of isolating,culturing and identifing in bladder smooth muscle cell of rabbit. Methods: Bladder smooth muscle cells were isolated from two adult male New Zealand White rabbits by collagenase digestion and cultured in DMEM medium supplemented with calf bovine serum. Morphology and expansion of the cells were observed. Cells were identified by smear of smear electron microscope and immunohistochemical methods(immunostaining with anti-α-actin antibodies). Results: The cells grew well and presented atypical morphological feature of smooth muscle cell with invert microscope(forming the 'hills and valleys'). Ninety-nine percent of the cells were smooth muscle cells identified by smear of smear,electron microscope and immunohistochemical methods.HE stain and immunostaining. Conclusion:This bladder smooth muscle cell of rabbit cultural method is convenience, fruitful and reliable.