猪膀胱平滑肌细胞的体外培养及鉴定

Culture and identification of porcine bladder smooth muscle cells in vitro

目的 探讨猪膀胱平滑肌细胞的体外分离、培养、增殖及鉴定的方法,为猪膀胱平滑肌细胞作为构建组织工程细胞的可行性提供实验依据.方法 选择4只滇南小耳猪,在无菌条件下切取猪膀胱组织,仔细分离膀胱平滑肌层,利用混合酶消化法获取膀胱平滑肌细胞,在DMEM/F12培养基中加入10%胎牛血清,观察细胞形态改变,生长及增殖过程,并利用免疫组织化学染色法鉴定细胞.结果 体外培养的细胞可融合成片,倒置相差显微镜下观察细胞为长梭形,并且表现出典型的"谷和峰"的形态,通过HE染色符合平滑肌细胞形态特征,平滑肌肌动蛋白染色呈阳性.结论 采用体外培养方法能够在体外大量培养出高纯度猪膀胱平滑肌细胞,猪膀胱平滑肌细胞可作为种子细胞用于构建组织工程膀胱.

Objective To explore the approaches of porcine bladder smooth muscle cells separation,culture,proliferation and identification in vitro,and provide evidence for tissue engineering bladder establishment.Methods The bladder were obtained from 4 Diannan small eared pigs,and the bladder smooth muscle cells were isolated by trypsin digestion and cultured by DMEM/F12 medium.Cell growth and proliferation were observed,and the bladder smooth muscle cells were identified by immunohistochemical staining.Results The cells cultured in vitro appeared confluent and were in long shuttle form with the typical morphology of “valley and peak”.These cells were in accord with the morphological characteristics of smooth muscle cells by HE staining,and they were positive for smooth muscle actin staining.Conclusions Smooth muscle cells of the porcine bladder are available through this approach which can be used in the construction of tissue engineered bladder.