人间充质干细胞免疫原性研究

目的探讨人间充质干细胞(hMSCs)特性及免疫原性,为进一步研究hMSCs移植特性提供实验依据。方法免疫组化方法鉴定hMSCs表面HLA-ABC、HLA-DR、CD80、CD86分子;流式细胞术检测其含量;RT-PCR检测HLA-ABC、HLA-DRmRNA基因片断;外周血淋巴细胞杀伤试验及 CCK-8比色法规察淋巴细胞增殖反应;将表达绿色荧光蛋白(GFP)的DNA复合物导入hMSCs进行大鼠脑内移植,观察hMSCs在脑内的存活情况。结果 hMSCs表面少量表达HLA-ABC分子,不表达 HLA-DR、CD80、CD86分子;有少量HLA-ABC mRNA基因片断存在,未发现HLA-DR mRNA基因片断;外周血淋巴细胞杀伤试验没有发现淋巴细胞增殖反应;大鼠脑内移植hMSCs一个月后仍可见有细胞存活。结论 hMSCs具有较弱的免疫原性。

Immunogenicity of human mesenchymal stem cells

Objective To explore the characters and immunogenicity of human mesenchymal stem cells (hMSCs) and to provide experimental data for future fundamental study on hMSCs transplantation. Methods Identification of the HLA-ABC, HLA-DR, CD80 and CD86 molecules on the surface of hMSCs was performed by using immunohistochemical methods and the cellular proportion was detected by using flow cytometry. RT-PCR was made to detect mRNA gene fragment of HLA-ABC and HLA-DR. Cytotoxicity of peripheral blood lymphocytes was assayed to make sure if hMSCs can make lymphoproliferation response. hMSCs were transplanted into rat brain to make sure if they can survive in the next 30 d. Results Only HLA-ABC molecules but not HLA-DR, CD80, CD86 molecules were expressed on the surface of hMSCs. HLA-ABC mRNA gene fragment was positively detected. Cytotoxicity assay of peripheral blood lymphocytes showed no lymphocyte proliferation. The transplanted hMSCs survived well in the rat brain one month later. Conclusion hMSCs possess weak immunogenicity.