腺病毒载体对骨髓间充质干细胞体外分化能力的影响  

Effect of adenovirus-mediated gene transfer on the differentiation potentials of bone marrow derived mesenchymal stem cells

作　　者：边素艳[1] 盖鲁粤[2] 叶平[1] 郭子宽[3,4] 王华[3,4] 胡泽斌[3,4] 

机构地区：[1]解放军总医院老年心血管二科,北京100853 [2]解放军总医院心血管内科,北京100853 [3]解放军总医院 [4]军事医学科学院放射医学研究所实验血液学研究室,北京100850

出　　处：《中国现代医学杂志》2010年第8期1152-1156,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金青年基金面上项目(No:30400182)

摘　　要：目的研究腺病毒载体对Wistar大鼠骨髓间充质干细胞体外分化能力的影响。方法密度梯度离心加贴壁培养法自2周龄Wistar大鼠骨髓中分离培养MSC,用携带绿色荧光蛋白基因的腺病毒载体(Ad-GFP)感染,48 h后用流式细胞仪检测感染效率;在特殊诱导剂诱导下,用碱性磷酸酶(ALP)染色、ALP活性及油红O染色分别鉴定腺病毒载体对MSCs成骨和成脂分化潜能的影响。结果 Ad-GFP感染MSC 48 h后97.82%的细胞表达GFP;MSC经特异性诱导剂诱导后可向脂肪及成骨细胞分化,Ad-GFP感染对MSC成脂及成骨分化能力无显著影响。结论腺病毒载体对大鼠骨髓MSC感染效率高,且不影响其体外多向分化潜能,是基因修饰MSC的理想载体。【Objective】 To identify whether adenovirus-mediated gene transfer strategy interferes with the differentiation potentials of rat bone marrow mesenchymal stem cells（MSC）.【Methods】 MSC were isolated from the femoral and tibial bones of 2-week-old Wistar rats by density gradient centrifugation and purified on the basis of their ability to adhere to plastic.MSC were infected by an adenoviral vector carrying green fluorescent protein gene（Ad-GFP） and 48 hours later,the infection efficiency was assessed with flow cytometry,and their osteogenic and adipogenic differentiation potentials were identified by histological and molecular methods.【Results】 Ad-GFP infected MSC with a high efficiency of 97.82% around.Gene-modified MSC could differentiate into adipocytes and osteoblasts in vitro under specific inductive conditions,which was indentified by Oil-Red O and alkaline phosphatase staining.Ad-GFP infection did not affect MSC in the osteogenic or adipogenic differentiation potential.【Conclusion】Exogenous gene transfer into Wistar rat’s MSC can be efficiently conducted by the adenoviral vector.Adenovirus-mediated gene transfer does not interfere with their differentiation capability,and this kind of adenoviral vectors could be used for the ex vivo modification of MSC.

关 键 词：大鼠 骨髓间充质干细胞 分化 

分 类 号：R331.22[医药卫生—人体生理学]