携带IL-18基因人脐带间充质干细胞的构建

目的构建携带白细胞介素-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工具。方法体外分离培养hUMSCs,流式细胞仪（FACS）检测hUMSCs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方法检测IL-1蛋白及mRNA表达水平。结果成功在体外分离和培养了hUMSCs,FACS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方法检测结果显示,IL-18基因转染至hUMSCs并能稳定表达。结论构建了携带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的实验工具。

CONSTRUCTION OF IL-18 GENE IN HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS

Objective To construct IL-18 gene in human umbilical cord mesenchymal stem cells for exploring a means of tumor targeted gene therapy. 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, and the CD34 and CD45 were negative, fitting the phenotype of hUMSCs. IL-18-hUMSCs was created successfully. RT-PCR and Western blot showed that IL 18 gene was transfected into hUMSCs and stably expressed. Conclusion hUMSCs that carries and stably expresses gene IL-18 has been created, which provides a new method for experimental research of targeted tumor gene therapy.