【关键词】　　膀胱移行上皮细胞　　细胞培养　　生物学特征　　犬中图分类号: Q 813　R318　　文献标识码: AEXPERI M ENTALSTUDYON CHARACTERIZATI ON OFNORMALCAN INEBLADDERTRANSITI ONALEPITHEL IAL CELLS CULTURED IN VITRO HA N P ing , YA N G Zhim ing , L I X iuqun, et al. D iv ision of S tem Celland T issue Eng ineering , S tate K ey L aboratory of B iotherapy , W est China H osp ital, S ichuan U niversity , ChengduS ichuan, 610041, P. R. China.E2m ail: H anp 2001a@ sohu. comCorresp ond ing author: YA N G Zhim ing , E2m ail: orthop @ p ublic. cd. cn【Abstract】　　Objective　To exp lore an effectivemethod to culture and purify canine bladder transitional ep ithelialcells. 　M ethods　B ladder tissue w as obtained from healthy puppy under sterile conditions.B ladder mucosa w asremoved from the remaining tissue w ith fine scissor and m inced into small p ieces, and then w ere dissociated into singlecell suspensions w ith 0. 125% tryp sin.The bladder ep ithelial cells w ere cultured in defined keratinocyte serum freemedium.The cells w ere passaged and purified by 01 05% tryp sin and 01 02% EDTA. M orphological characterizationw ere studied under inverted phase contrast m icroscope and transm ission electron m icroscope. Exp ression of cell specificmarker p rotein w as assessed by immunohistochem istry. 　Results　Canine bladder transitional ep ithelial cells could beefficiently cultivated and expanded in serum2free medium w ithout fibroblast contam ination. The cells could be passaged426 times w ithout a distinguished decrease in cell p roliferation.The cells w ere characterized by w ell2developed m icro2filament and desmosome junction under transm ission electron m icroscope.I mmunohistochem ical staining w ith broadlyreacting anti2cytokeratin antibodies (A E1 A E3) confirmed the ep ithelial phenotype of the cells. D ifferent generations ofcells show ed dip loid cells. 　Conclusion　A large number of bladder transitional ep ithelial cells can be obtained fromsmall bladder tissue w ith our digestion method.The cultured bladder ep ithelial cells can be p roliferated to sufficientquantities for further reconstructive purposes.【Key words】　　B ladder transitional ep ithelial cell　　Cell culture　　B iological characterization　　CanineFoundation item: Postdoctoral Science Foundation of China(20070410389)　　组织工程技术的发展为泌尿道修复重建开辟了崭新的途径, 其基本技术路线包括种子细胞的培养、 支架材料的研制、 细胞与支架材料的相互作用以及工程化基金项目: 中国博士后科学基金资助项目 (20070410389)