摘要
目的探讨慢病毒载体用于羊骨髓间充质干细胞(mesenchymal stem cells,MSCs)基因表达关键问题。方法携带绿色荧光蛋白基因(Zs Green)的慢病毒载体感染羊MSCs获得慢病毒转染MSCs(MSCz),采用流式细胞仪、MTT检测、三系分化等方法对MSCz比例、细胞形态、增殖及分化潜能进行评价。结果 MSCz保持骨髓间充质干细胞特有的长梭形细胞形态,并高效表达绿色荧光蛋白。流式细胞仪分析显示MSCz比例随感染复数(multiplicity of infection,MOI)增加而升高,在MOI值为20时MSCz比例可达71.2%。细胞生长曲线分析表明MSCz与MSCs增殖模式和能力相同。三系分化证实MSCz与MSCs在向骨、脂肪和软骨细胞分化方面没有差异。过氧化氢和低氧处理表明MSCz、MSCs对这些不利环境反应相同。结论慢病毒载体是羊MSCs基因表达的良好载体。
Objective To study the feasibility of lentiviral vector-mediated modification on ovine marrow mesenchymal stem cells（MSCs） for gene expression. Methods Ovine MSCs were transduced into lentiviral vector MSCz by Zs Green lentiviral vector, and the effects of transduction on morphology, proliferation and differentiation potential were evaluated using flow cytometry, MTT assay and trilealinige differentiation, respectively. Results MSCz maintained the specific morphology of mesenchymal stem cells, and expressed Zs Green proteins as detected by fluorescent microscopy. The percentage of MSCz increased with the rise of MOI based on flow cytometry analysis, and it could reach 71.2% with MOI of 20. MTT assay showed that the proliferation ability of MSCz were same as MSCs. The multipotent ability of MSCz showed no difference in differentiation towards adipose, chondrocyte and osteoblast from MSCs. Stress by hydrogen peroxide or culture under hypoxia showed that MSCs and MSCz had the same response to these hostile environments. Conclusion Lentiviral vector is a good vector for transducing ovine MSCs for gene expression.
作者
梁文涛
丁毅
王嵘
张勇
李则学
梁凯
赵云山
LIANG Wentao DING Yi WANG Rong ZHANG Yong LI Zexue LIANG Kai ZHAO Yunshan(Institute of General Surgery, Chinese PLA General Hospital, Beijing 100853, China Institute of Anorectal Surgery, the General Hospital of the Chinese People＇s Armed Police Forces, Beijing 100039, China Department of Cardiovascular Surgery, Chinese PLA General Hospital, Beijing 100853, China)
出处
《解放军医学院学报》
CAS
2016年第12期1284-1288,共5页
Academic Journal of Chinese PLA Medical School
关键词
羊骨髓间充质干细胞
慢病毒载体
转染
ovine marrow mesenchymal stem cells
lentiviral vector
transfection
