两种体外培养兔膀胱变移上皮细胞方法的比较

Comparison between the two kind methods of culture in vitro for vesical transitional cell in rabbits

目的比较组织贴块法和酶消化法体外培养兔膀胱变移上皮细胞的优缺点,探索简单、高效培养膀胱变移上皮细胞的方法,为膀胱组织工程种子细胞的培养提供实验基础。方法采用组织贴块法和酶消化进行兔膀胱变移上皮细胞体外培养,相差显微镜下观察其生长特点,应用细胞免疫荧光染色对细胞进行鉴定,测定细胞生长曲线了解细胞生长的基本规律。结果培养的细胞具有上皮细胞的典型形态,生长曲线呈"S"形,角蛋白(CKAE1/AE3)单克隆抗体免疫荧光染色阳性,波形蛋白染色阴性。结论组织贴块法和酶消化法均能成功培养出兔膀胱变移上皮细胞。组织贴块法原代培养时间长,但是成功率高。酶消化法可以在较短时间内获取大量细胞,但是操作步骤多,细胞容易污染或损伤。

Objective Bladder transitional epithelial cells from rabbit were primarily cultured by tissue explant and enzyme digestion method respectively to explore the better culture technique in vitro and provide seed cells for engineering of bladder tissue.Methods Rabbit bladder transitional epithelial cells were primarily cultured by tissue explant and enzyme digestion method respectively.The cell morphology and proliferation characteristics were oberserved.The grow curve of the cells was described through cell counting.The cells were identified by cell morphology and immunofluorescence.Results The bladder transitional epithelial cells were obtained by the two methods possessed a typical morphological feature of epithelial cells.And the morphology and growth characteristics were essentially consistent.The growth curve of the cells was similar to 'S' shape.It showed vimentin negative and cytokeratin AE1 /AE3 positive.Conclusion Both the culture methods could obtain high purity of bladder transitional epithelial cells.The method of tissue explant has a higher success rate but spending more time than that of enzymatic digestion.