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].现代生物医学进展英文版,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 |
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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) |
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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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