兔正常膀胱移行上皮细胞的体外培养和鉴定

目的：摸索膀胱移行上皮细胞的合适培养条件，为建立满足组织工程需要的尿路上皮细胞体培养体系提供实验基础。方法：采用无血清培养系统，以组织块法原代培养兔正常膀胱移行上皮细胞并传代。动态观察细胞形态变化和生长增殖状况，进行免疫细胞化学染色鉴定细胞来源。结果：原代第4开始有上皮样细胞自组织块边缘长出，第10-14天汇合，呈典型铺路石样外观。2代超细胞生长4-7d后汇合，未发现成纤维样细胞混杂生长。细胞可传至6代以上。细胞角蛋白AE1/AE3单抗染色各代细胞均呈阳性反应。结论：分离培养的是单一的膀胱移行上皮细胞，细胞具有一定的增殖传代能力。

Culture and Characterization of Normal Rabbit Bladder Transitional Epithelial Cells in Vitro

Objective: To explore suitable culture conditions of bladder transitional epithelial cells for further research on establishing a promising culture method of urothelial cells that meets the requirements of tissue engineering.Methods: The primary culture of normal rabbit bladder transitional epithelial cells were initiated by using explant culture method and the cells were subcultured in serum-free media. Morphological and proliferative changes of cells were observed dynamically.Cell specificity was confirmed by cytokeratin analysis.Results:The outgrowth of epithelial-like cells from bladder mucosa explants occurred within 4 days in primary culture.After 6 to 10 days monolayers attained confluence with a typical cobble like appearance.Subsequent subcultures achieved confluence winthin 4 to 7 days and a consistently high rate of growth was sustained for at least 6 passages. Positive staining of immunocytochemistry with monoclonal antibody against cytokeratin AE1/AE3 was detected in all passages of cells.Conclusion:The cultured cells were exclusively bladder transitional epithelial origin and retained the capacity of differentiation and proliferation.