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脂肪干细胞原代培养及诱导成肌细胞的实验研究
引用本文:宋小飞,傅强,徐月敏,崔磊.脂肪干细胞原代培养及诱导成肌细胞的实验研究[J].上海交通大学学报(医学版),2009,29(5).
作者姓名:宋小飞  傅强  徐月敏  崔磊
作者单位:宋小飞,傅强,徐月敏,SONG Xiao-fei,FU Qiang,XU Yue-min(上海交通大学,第六人民医院泌尿外科,上海,200233);崔磊,CUI Lei(上海市组织工程研究与开发中心,上海,200230)  
基金项目:上海市科委临床重点课题基金 
摘    要:目的 探讨大鼠脂肪干细胞(ADSC)的培养及诱导分化为成肌细胞的方法 .方法 取SD大鼠脂肪组织,酶消化法分离、培养ADSC,免疫荧光和流式细胞仪检测相关表面抗原CD90、CD105和CD34的表达.取培养至第2代处于对数生长期细胞,分别用含5-氮杂胞苷(5-aza)的诱导培养液(诱导组)和基础培养液(对照组)进行培养.诱导时间为7、14、21、28和35 d,倒置相差显微镜观察细胞的生长情况和形态变化,免疫荧光和流式细胞仪检测成肌细胞特异性抗原desmin和myosin的表达.结果 成功从大鼠脂肪组织分离、培养出ADSC,相关表面抗原表达检测证实其干细胞特性.诱导组细胞诱导28 d后呈现成肌细胞特有的"漩涡"样生长形态,单个细胞表现出多核化;免疫荧光和流式细胞仪检测显示,desmin和myosin的表达率在诱导28 d时达最高,分别为52.57%和50.04%,而诱导前和对照组细胞均呈阴性表达.结论 成功从大鼠脂肪组织分离、培养ADSC,含5-aza的诱导培养液可将其诱导分化为成肌细胞,诱导 28 d成肌细胞特异性抗原表达率最高.

关 键 词:脂肪干细胞  细胞分化  成肌细胞  压力性尿失禁  5-氮杂胞苷

Primary culture of adipose-derived stem cells and differentiation induction into myoblasts
SONG Xiao-fei,FU Qiang,XU Yue-min,CUI Lei.Primary culture of adipose-derived stem cells and differentiation induction into myoblasts[J].Journal of Shanghai Jiaotong University:Medical Science,2009,29(5).
Authors:SONG Xiao-fei  FU Qiang  XU Yue-min  CUI Lei
Institution:1.Department of Urology;The Sixth People's Hospital;Shanghai Jiaotong University;Shanghai 200233;China;2.Shanghai Tissue Engineering Research and Development Center;Shanghai 200230;China
Abstract:Objective To explore the method of culture of rat adipose-derived stem cells(ADSCs) and differentiation induction into myoblasts. Methods Adipose tissues were obtained from SD rats,and were isolated by enzyme digestion and cultured into ADSCs.The expression of surface antigen CD90,CD105 and CD34 was detected by immunofluorescence and flow cytometry.ADSCs of the second passage with logarithmic growth were obtained,and culture media containing 5-azacytidine(5-aza) and basic culture media were employed for cel...
Keywords:adipose-derived stem cell  cell differentiation  myoblast  stress urinary incontinence  5-azacytidine  
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