人脂肪干细胞与猪尿路上皮细胞隔离共培养后向尿路上皮样细胞分化

Adipose-derived stem cells can be induced into urothelium-like phenotype in vitro by indirect co-culture with porcine urothelium cells

目的研究人脂肪干细胞与猪尿路上皮细胞体外隔离共培养后向尿路上皮样细胞转化的可行性。方法从人抽脂术后脂肪组织中提取脂肪干细胞,同时从猪膀胱中获取尿路上皮细胞,两种细胞体外培养扩增后,通过"Trans-well"体系进行体外隔离共培养3周。倒置显微镜下观察脂肪干细胞形态变化。RT-RCP鉴定诱导后人脂肪干细胞中尿路上皮特异性标记Uroplakin-Ⅱ在基因水平的表达,免疫荧光鉴定AE1/AE3和Uroplakin-Ⅱ在蛋白水平的表达情况。结果人脂肪干细胞与猪尿路上皮细胞体外隔离共培养3周后,倒置显微镜可见细胞折光性变强,细胞触角缩短,依然保持良好的增殖状态,但未见典型的尿路上皮细胞"铺路石样"改变。免疫荧光发现AE1/AE3和Uroplakin-Ⅱ均有表达,RT-PCR检测发现Uroplakin-Ⅱ基因表达阳性。结论人脂肪干细胞与猪尿路上皮细胞隔离共培养后,可以表达成熟尿路上皮特异性标记,说明脂肪干细胞可以向尿路上皮样细胞分化。

Objective To explore the feasibility of phenotype transformation of human adipose-derived stem cells（ ADSCs）towards bladder urothelium cells by co-culture with porcine urothelium through＂ Trans-well＂ system. Methods ADSCs were isolated from human subcutaneous adipose tissue and co-cultured with pig urothelium through＂ Trans-well ＂ system. The specific marker expressions of urothelium in ADSCs were examined by immunohistochemistry and RT-PCR at 3 weeks to evaluate the potential phenotype transformation ,and the cell shape was observed at the same time. Results The cell shape was partially changed from a fibroblast-like cell to a polygonal cell in 3 weeks. But we can＇t find the ＂paving stone ＂cell shape which is specific of urothelium. After 3 weeks of culture ,ADSCs expressed AE1/AE3 at protein level, Uroplakin-Ⅱ at both protein and mRNA level. But the expression of the two makers was weaker than that in urothelium. Conclusions Our findings reveal that ADSCs can be induced into urothelium-like phenotype by indirect co-culture with pig urothelium.