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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。
Objective: To construct human umbilical cord mesenchymal stem cells (hUMSCs) carrying interleukin-18 (IL-18) gene and to provide a tool for tumor-targeted gene therapy. Methods: hUMSCs were isolated and cultured in vitro. The cellular immunophenotype of hUMSCs was detected by flow cytometry (FACS). The lentivirus expressing IL-18 gene was transfected into hUMSCs by gene recombination technique. The expression of IL-18 protein and mRNA was detected by RT-PCR and Western blot. Results: hUMSCs were successfully isolated and cultured in vitro. The results of flow cytometry showed that hUMSCs expressed CD29, CD44 and CD105 but not CD34 and CD45, which was in line with the phenotype of hUMSCs. The hUMSCs carrying IL-18 gene were successfully constructed. The results of RT-PCR and Western blot indicated that IL-18 gene was transfected into hUMSCs and stably expressed. CONCLUSION: The construction of hUMSCs carrying IL-18 gene and stable expression of IL-18 provide a new experimental tool for the experimental study of tumor-targeted gene therapy.