小鼠同种异体骨髓间充质干细胞的免疫原性及其存活分化研究

目的研究小鼠同种异体骨髓间充质干细胞（MSCs）的免疫生物特性及其在宿主肌袋内的存活和分化能力.方法分离培养GFP转基因FVBN小鼠的MSCs,定量分别将其注入6组（植入后1 d、1周、2周、3周、5周、7周）FVBN小鼠的左腿肌袋内.流式细胞仪检测注射后各时间段宿主外周血中调节性T细胞（CD4＋CD25＋Tcel）l的变化规律来推断同种异体MSCs的免疫原性,应用荧光示踪,免疫组化等方法研究同种异体MSCs在小鼠肌袋内的存活和分化能力.结果成功培养出小鼠MSCs,宿主调节性T细胞的比例在植入后各时间段之间以及和对照组相比无统计学意义（P〉0.05）.在1周内的宿主肌袋中可观察到MSCs,但并未向成软骨分化,之后无法检测到MSCs.结论小鼠同种异体MSCs植入到肌袋后,可能具有低免疫原性.在局部无法长期观察同种异体MSCs及其分化,原因可能为细胞死亡或迁移,以及局部缺乏相应诱导成软骨的微环境.

Immunogenicity, Surviving and Differentiation of Mouse's Mesenchymal Stem Cells in Host's Muscle after Allograft

Objectives To study the immunogenicity,surviving and differentiating ability of MSCs after allograft.Methods We separated and cultured the GFP transinfected MSCs of FVBN mouse,then we injected the MSCs into the tibia muscles of FVBN mice in 6 groups（1 d,1 week,2 weeks,3 weeks,5 weeks,and 7 weeks after implantation）.The changes in the ratio of T regular cell（CD4＋CD25＋Tcell）in host＇s peripheral blood were detected by flow cytometry to deduce the immunogenicity of allograft MSCs.The surviving and differentiating ability of allograft MSCs in the tibia muscles of mice was detected by immuno-histochemistry staining.Results MSCs of mice were cultured successfully.There was no significant difference in the T regular cell＇s ratio among different experimental and control groups（P 0.05）.The MSCs were observed just in one week after allograft,but they had no tendency of chondrogenic differentiation,and no MSCs were found later.Conclusions Allograft MSCs has low immunogenicity after implantation in muscles of mice.Allograft MSCs and their differentiation can not be observed in long term,this may be because of death or migration of cells,and microenviroment without inducing factors.