慢病毒介导髓细胞触发受体2过表达质粒转染促进人骨髓间充质干细胞向神经细胞分化  

Lentivirus-mediated over-expression plasmid transfection of myelocyte trigger receptor 2 promotes bone marrow mesenchymal stem cells to neurons

作　　者：刘峰 李晓飞[2] 刘艳[2] LIU Feng;LI Xiao-fei;LIU Yan(Department of Neurosurgery,Dezhou Traditional Chinese Medicine Hospital,Dezhou 253000;Laoshan Hospital Operating Room,Qingdao University Affiliated Hospital,Qingdao 266003,China)

机构地区：[1]山东省德州市中医院神经外科,山东德州253000 [2]青岛大学附属医院崂山院区手术室,山东青岛266003

出　　处：《解剖科学进展》2020年第5期570-574,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81672197)。

摘　　要：目的探究慢病毒介导TREM-2质粒转染促进骨髓间充质干细胞向神经细胞转化的研究。方法利用基因过表达技术,构建TREM-2过表达载体,慢病毒转染骨髓间充质干细胞(BMSCs),利用流式细胞仪检测BMSCs的表面蛋白,利用RT-PCR和Western blot检测神经细胞表面蛋白胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和神经元特异性烯醇化酶(neuron-specific enolase,NSE)以及神经丝蛋白(neurofilament protein,NF),巢蛋白的表达。结果在第2代BMSCs,表面蛋白CD29阳性率为98.03%,CD90阳性率为96.92%,CD45阴性率为77.2%,共表达CD29、CD90且CD45表达阴性的细胞数占总细胞数的96.89%。BMSCs细胞表面CD34和CD44免疫组织化学染色呈阳性。慢病毒转染效率为90%以上,转染组中GFAP-,NSE-,NF-和巢蛋白mRNA和蛋白的表达量明显高于空白对照组和阴性对照组。结论慢病毒介导TREM-2过表达载体转染促进骨髓间充质干细胞BMSCs向神经细胞转化,为中枢神经损伤和脊髓损伤的治疗提供种子细胞。Objective To investigate the effect of lentivirus-mediated TREM-2 plasmid transfection on the transformation of bone marrow mesenchymal stem cells into neurons.Methods TREM-2 overexpression vector was constructed by gene overexpression technology,lentivirus was transfected into bone marrow mesenchymal stem cells(BMSCs).The surface proteins of BMSCs were detected by flow cytometry,the expressions of neuron-specific enolase(NSE),neurofilament protein(NF)and glial fibrillary acidic protein(GFAP)and nestin were detected by RT-PCR and Westernblot.Results The positive rate of CD29 was 98.03%,CD90 was 96.92%,and the negative rate of CD45 was 77.2%,the total number of cells expressing CD29,CD90 and CD45 was 96.89%in the second generation BMSCs,with CD34-and CD44-positive on the surface of BMSCs by immunohistochemical staining.The transfection efficiency of lentivirus was over 90%,the expression levels of GFAP-,NSE-,NF-and nestin-mRNA and proteins were significantly higher in transfection group than in the blank control or negative control.Conclusion Lentivirus-mediated TREM-2 overexpression vector transfection promotes the transformation of bone marrow mesenchymal stem cells into neurons,which would provide seed cells for repair of nervous system injury.

关 键 词：髓细胞触发受体2 人骨髓间充质干细胞 慢病毒转染 胶质纤维酸性蛋白 神经元特异性烯醇化酶 

分 类 号：Q813.1[生物学—生物工程]