人体尿道粘膜上皮细胞的连续培养

目的：探讨人尿道粘膜上皮细胞体外培养技术，为构建组织工程化尿道提供种子细胞。方法：取尿道下裂患者尿道粘膜，经中性蛋白酶和胰蛋白酶消化，获取粘膜上皮细胞接种于含8％胎牛血清培养基中连续培养，观察细胞形态变化及生、增殖过程。结果：体外培养细胞长满后呈上皮细胞特有的铺路石样外观，体外可连续传9-10代，生长期50-60d。细胞角质蛋白免疫组织化学染色阳性，透射电镜下可见上皮细胞间特有的桥粒结构。结论：尿道粘膜上皮细胞可在体外连续培养，4代以内的培养细胞可用于组织工程化尿道的构建。

Serial cultivation of human urethral epithelium

Objective To establish a method of cult ur ing human urethral epithelium.Methods Specimens from urth ral epithelium of hypospadias were dissociated into single cell suspension by di spase and trypsin.The cells were grown in medium with 8 % fetal bovine serum to observe morphological characteristics.Results The cells c ould be maintained in culture up to 9-10 passages with a survival of 50 to 60 days.Electron microscopy revealed desmosomes between the culture cells.The cells showed positive staining for cytokeratin antibody.Conclusion The culture technique with fetal bovine serum for growing urethral epitheli um has been established.The culturing cells of 4 passages could be used in uret hral contruction of tissue engineering.