天津医药

• 实验研究 •    下一篇

大鼠脂肪间充质干细胞的分化潜能鉴定及XIAP基因修饰

白耀邦,李博,陈庆良,姜楠,陈晓波   

  1. 天津市胸科医院心外科
  • 收稿日期:2012-10-16 修回日期:2013-03-13 出版日期:2013-08-15 发布日期:2013-08-15
  • 通讯作者: 白耀邦

Preparation and Characterization of Rat Adipose-Derived Stem Cells and XIAP Modified Adipose-Derived Stem Cells

BAI Yaobang ,LI bo ,CHEN Qingliang ,JIANG Nan ,CHEN Xiaobo   

  1. Tianjin Chest Hospital, Tianjin 300051, China
  • Received:2012-10-16 Revised:2013-03-13 Published:2013-08-15 Online:2013-08-15
  • Contact: BAI Yaobang

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摘要： 【摘要】 目的 分离培养扩增大鼠脂肪源间充质干细胞(ADSCs)，以活体标记并鉴定其分化潜能，对比XIAP修饰的细胞与未处理组细胞的抗凋亡能力。方法 无菌条件下取大鼠一侧腹股沟脂肪组织，Ⅰ型胶原酶消化法分离培养脂肪间充质干细胞，胰酶消化法传代扩增。检测细胞分化为脂肪细胞、软骨细胞及成骨细胞潜能，转染XIAP表达质粒进入ADSCs，通过Western blotting等方法检测XIAP蛋白的表达及抗凋亡能力。结果  脂肪间充质干细胞呈长梭形漩涡样生长，细胞流式鉴定显示CD29、 CD44、 CD90、 CD105均呈高表达，并在特定诱导剂下分化为脂肪细胞、软骨细胞或成骨细胞。利用过表达XIAP的ADSCs可以增强ADSCs的抗凋亡能力，提高其耐受缺血、缺氧能力,增强其功能。结论   来源于大鼠腹股沟脂肪分离获得的脂肪源干细胞易于培养和传代扩增，并可活体标记，并具有多向分化潜能，可作为组织工程的种子细胞。

关键词: 间质干细胞, 脂肪组织, X连锁凋亡抑制蛋白质, 软骨细胞, 成骨细胞, 细胞分化, 大鼠, Wistar

Abstract: Objective   To isolate and cultivate rat ADSCs in vitro, and to identify its differentiation potential. Modification of ADSC and test test the antiapoptotic ability after XIAP modification. Methods   ADSCs were isolated from rat groin fat pads by CollagenaseⅠdigestion under sterile condition. ADSCs were passaged and amplified with 10% FBS DMEM. The multi-differentiation potential of ADSCs were verified by cultivated with differentiation medium. Electrotransduction of XIAP expression plasmid into stem cells was done followed with normal procedure. The antiapoptotic effection of XIAP transduction was verified by the downregulation of caspase activation.Results   In vitro cultured ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. ADSCs were positive expression of CD29、CD44、CD105 and CD90, ADSCs can differentiate into lipocytes,chondrocyte and osteoblasts under certain conditions. The over-expression of XIAP in ADSCs can enhance the ADSCs antiapoptotic ability, improve its tolerance of ischemia and hypoxia.Conclusion   ADSCs were isolated from rat subcutaneous fat pads and were easily cultivated, passaged and amplified. ADSCs were highly effective modified by electrotransduction, and differentiated into osteoblasts、chondrocyte and adipocytes under certain conditions. ADSC is a better resource for being used in cell therapy and tissue engineering.

Key words: mesenchymal stem cells, Adipose tissue, X-linked inhibitor of apoptosis protein, chondrocytes, osteoblast, cell differentiation, rats, Wistar