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。
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.
出处
《现代生物医学进展》
CAS
2014年第25期4847-4851,共5页
Progress in Modern Biomedicine
基金
Natural Science Foundation of China(81302290)
Natural Science Foundation of Shandong(ZR2013HM019)~~
关键词
白介素-18
人脐带间充质干细胞
基因治疗
Interleukin-18
Human umbilical cord mesenchymal stem cells
Gene therapy
