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

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。

Construction of IL-18 Gene in Human Umbilical Cord Mesenchymal Stem Cells and Its Significance

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 success- fully 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.