兔膀胱移行上皮细胞的培养和鉴定

The culture and identification of rabbit bladder transitional epithelial cells in vitro

目的建立一种有效、简便的兔膀胱上皮细胞体外原代培养方法。方法以DMEM：F12为基础培养基，添加10％胎牛血清（fetal bovine serum，FCS）、表皮细胞生长因子（epidermal growth factor，EGF）等营养成分，在37℃、5％CO2的孵箱内培养，观察细胞形态变化及生长、增殖过程，并进行免疫荧光鉴定。结果细胞在第10～14d融合形成单层，细胞形态呈多角形的铺路石子状，经免疫荧光鉴定证实为移行上皮细胞。结论此培养方法能在短时间内获得大量移行上皮细胞，为进一步采用组织工程技术构建尿道提供种子细胞，为治疗重度尿道下裂、尿道下裂残废和较长的后尿道狭窄等顽症提供依据。

Objective To establish a culture method of rabbit bladder transitional epithelial cells in vitro. Methods Separated bladder transitional epithelial cells were cultured in DMEM： F12（1：1） supplemented with nutrition including 10%fetal bovine serum, EGF, and so on. Cells were incubated at 37℃ in a humidified atmosphere of 5% CO2. Morphology of the cells was observed. Urothelial cell specific protein （cytokeratin） was identified by irnmunofluorescence. Results After 10-14 days of culture, confluent growth was noted. Positive staining of immunocytochemistry with monoclonal antibody against cytokeratin was detected in all passages of cells. Conclusions The culture technique for growing urothelial cell has been established. The culturing cells may be used in urethral construction of tissue engineering and then be used to treat severe hypospadias, hypospadias cripples and urethral stricture in the future.