兔膀胱移行上皮种子细胞的培养及扩增

In vitro amplification of the bladder transitional epithelial cells

目的 对膀胱移行上皮细胞进行原代及传代培养 ,探讨用于膀胱组织工程研究的移行上皮种子细胞的培养方法。方法 采用组织块培养法 ,将获取的移行上皮组织小块在添加附加成分的无血清F12培养基进行培养 ,观察培养细胞的形态及生长增殖情况 ,并对传代培养的细胞进行免疫组化染色鉴定。结果 原代移行上皮组织培养约 9～ 10d时细胞可达 90 %的汇合 ,传代培养的移行上皮细胞生长稳定期较长 ,细胞至 5～ 6代时生长状态仍然良好。免疫组化染色显示培养细胞为移行上皮细胞。结论 采用组织块培养法能成功地原代培养兔膀胱移行上皮细胞 ,在添加附加成分的F12培养基中移行上皮细胞能够稳定地传代培养 ,表明该法可为膀胱组织工程的研究提供一定数量活性良好的移行上皮细胞。

Objective The bladder transitional epithelial cells were primary cultured?passaged and extensively expanded. To investigate the way for acquiring large quantites of well differentiated urothlial cells which will be used for bladder tissue engineering.Methods The bladder transitional epithelial explants were cultured in serum free F12 medium. Form and kinetic growth of the urothlial cells were observed, cytological immunocytochemical analysis against broadly reacting anti cytokeratin antibodyies(AE1/AE3) was also performed.Results The results showed the cells possessed a stable proliferation from the second passage to the sixth passage, implying that this method is ideal for the primary culture and subculture of the urothelium cells. Primary cells conflunce attached to 90% in the ninth or tenth day. Transitional epithelial cells were stained positively for broadly reacting anti cytokeratin antibodyies(AE1/AE3).Conclusion The bladder transitional epithelial cells can be successfully cultured by primary explants culture. In serum free F12 medium, urothlial cells can be cultured and effected expanded.This method of explants cultures will be of benefit for the bladder tissue engineering.