hPDGF-A／hBD2双基因转染对大鼠骨髓间充质干细胞生物学特性的影响  

Effects of hPDGF-A/hBD_2 genes transfection on rat bone marrow mesenchymal stem cells

作　　者：冯一梅[1] 徐辉[1] 邹仲敏[1] 郝磊[1] 王涛[1] 邓均[1] 董世武[1] 粟永萍[1] 程天民[1] 

机构地区：[1]第三军医大学军事预防医学院防原医学教研室,全军复合伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆400038

出　　处：《第三军医大学学报》2008年第6期472-476,共5页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划项目(“973”项目)(2005CB522605);全军医学科研“十一五”科技攻关项目(06G076)~~

摘　　要：目的探讨hPDGF-A/hBD2腺病毒双基因转染大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)后,对BMSCs本身生物学特性的影响。方法体外分离培养大鼠BMSCs,应用GFP标记的腺病毒表达载体系统Adv-hPDGF-A-IRES-hBD2,以脂质体法转染293细胞,再以病毒上清液感染BMSCs,对转染目的基因后的BMSCs进行形态学观察,绘制生长曲线,测定其细胞周期以及向成脂、成骨、成肌诱导分化和鉴定。结果hPDGF-A/hBD2双基因成功转入BMSCs后,未发现其对细胞活性有明显作用,基因修饰后的BMSCs仍具有成体干细胞所具备的增殖能力,以及向成脂、成骨、成肌等方向分化的潜能。结论BMSCs是重组腺病毒转染的良好靶细胞,hPDGF-A/hBD2基因修饰的BMSCs仍可作为满意的细胞治疗的种子细胞。Objective To study the biological characteristics of platelet-derived growth factor A and human beta defensin 2 （hPDGF-A/hBD2） gene-modified rat bone marrow mesenchymal stem eells （BMSCs）. Methods By using liposome transfection technique, recombinant adenovirus vector expressing hPDGF-A/ hBD2 （Adv-hPDGF-A-IRES-hBD2 ） labeled with GFP was transfected into 293T cells for virus packaging and amplification. BMSCs were isolated, cultured and infected by adenovirus-containing supernatant. The exogenous gene-modified BMSCs were comprehensively studied on their biological features, in terms of morphology, cell growth curve, cell cycle, and adipogenic, osteogenic and myogenic differentiation ability. Results hPDGF-A-IRES-hBD2 gene-modified BMSCs did not show obvious changes in cell viability, proliferation, cell cycle distribution or cell differentiation. Conclusion BMSCs were not only good carriers for exogenous hPDGF-A and hBD2 genes but also seed cells for cell therapy even after hPDGF-A/hBD2 modification.

关 键 词：骨髓间充质干细胞 基因转染 细胞生物学特性 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]