文章摘要
王琳 李福年 柳晓义 姜丹丹 吕志栋 曲慧利.携带白介素-18 基因的人脐带间充质干细胞的构建及其意义[J].,2014,14(25):4847-4851
携带白介素-18 基因的人脐带间充质干细胞的构建及其意义
Construction of IL-18 Gene in Human Umbilical Cord Mesenchymal StemCells and Its Significance
  
DOI:
中文关键词: 白介素-18  人脐带间充质干细胞  基因治疗
英文关键词: Interleukin-18  Human umbilical cord mesenchymal stemcells  Gene therapy
基金项目:国家自然科学基金项目(81302290);山东省自然科学基金项目(ZR2013HM019)
作者单位
王琳 李福年 柳晓义 姜丹丹 吕志栋 曲慧利 青岛大学医学院附属医院乳腺外科 
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中文摘要:
      目的:构建携带白介素-18(IL-18)基因人脐带间充质干细胞(hUMSCs),为肿瘤靶向性基因治疗研究提供一种工具。方法: 体外分离培养hUMSCs,流式细胞仪(FACS)检测hUMSCs 的细胞免疫表型。应用基因重组技术将表达IL-18 基因的慢病毒转染 至hUMSCs,利用RT-PCR 及Western blot 法检测IL-18 的蛋白及mRNA表达水平。结果:成功在体外分离和培养了hUMSCs,流 式细胞仪检测结果显示hUMSCs 表达CD29、CD44 和CD105,而不表达CD34 和CD45, 符合hUMSCs 的表型。成功构建携带 IL-18 基因的hUMSCs,RT-PCR 及Western blot法检测结果提示IL-18基因转染至hUMSCs并能稳定表达。结论:构建携带IL-18 基因的hUMSCs并稳定表达IL-18,为肿瘤靶向性基因治疗实验性研究提供了一种新实验工具。
英文摘要:
      Objective:To construct IL-18 gene in human umbilical cord mesenchymal stem cells for exploring a means of tumor targeted gene therapy research.Methods:hUMSCs were isolated and cultured in vitro. The immunophenotype of hUMSCs was detected by flow eytometry. The lentivirus vector containing human IL-18 gene was constructed and transfected into hUMSCs. The IL-18 mRNA and protein expression level was detected by semi-quantitative RT-PCR and Western blot.Results:hUMSCs were isolated and cultured successfully. The FACS indicated that CD29,CD44 and CD105 were positive; However the CD34 and CD45 were negative. So it med the phenotype of hUMSCs. IL-18-hUMSCs was produced successfully. RT-PCR and Western blot showed that IL-18 gene was successfully transfected into hUMSCs and stable expressed.Conclusion:Lentivirus-IL-18 vector can be transfected and stable expressed in hUMSCs that it provided a new experimental method of tumor targeted gene therapy experimental study.
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