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(英文)  

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

作　　者：王琳[1] 李福年[1] 柳晓义[1] 姜丹丹[1] 吕志栋[1] 曲慧利[1] 

机构地区：[1]青岛大学医学院附属医院乳腺外科,山东青岛266003

出　　处：《现代生物医学进展》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)~~

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.

关 键 词：白介素-18 人脐带间充质干细胞 基因治疗 

分 类 号：R392.11[医药卫生—免疫学]