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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
  • ISSN号:1674-0785
  • 期刊名称:中华临床医师杂志(电子版)
  • 时间:2012
  • 页码:2099-2102
  • 分类:R687[医药卫生—骨科学;医药卫生—临床医学;医药卫生—外科学]
  • 作者机构:[1]上海交通大学医学院附属第九人民医院泌尿外科,200011
  • 相关基金:国家自然科学基金面上项目(81070605);上海交通大学医工交叉基金面上项目(YG2011MSl4)
  • 相关项目:脂肪来源干细胞诱导尿路上皮细胞及其机制的研究
中文摘要:

目的研究人脂肪于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巾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AEl/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AEl/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。

英文摘要:

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.

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期刊信息
  • 《中华临床医师杂志(电子版)》
  • 主管单位:
  • 主办单位:中华医学会
  • 主编:
  • 地址:北京市西直门北大街32号B座606C室
  • 邮编:100082
  • 邮箱:lcdoctor@163.com
  • 电话:010-62219211
  • 国际标准刊号:ISSN:1674-0785
  • 国内统一刊号:ISSN:11-9147/R
  • 邮发代号:
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引
  • 被引量:30430