人脐带间充质干细胞分化为胰岛素分泌细胞的免疫原性变化  

Immunogenicity of insulin producing cells differentiating from human umbilical cord mesenchymal stem cells

作　　者：李兰兰 李宁 杨晓菲[2] 陈桃[2] 欧刚卫[1] 李富荣[2] Li Lan-lan;Li Ning;Yang Xiao-fei;Chen Tao;Ou Gang-wei;Li Fu-rong(Zunyi Medical University, Zunyi 563000, Guizhou Province, China;Translational Medicine Collaborative Innovation Center, the Second Clinical Medical College （Shenzhen People’s Hospital）, Jinan University, Shenzhen 518020, Guangdong Province, China)

机构地区：[1]遵义医学院,贵州省遵义市563000 [2]暨南大学第二临床医学院转化医学协同创新中心,广东省深圳市518020

出　　处：《中国组织工程研究》2018年第13期2045-2050,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81670702;81270857);广东省自然科学基金(2017030310646);深圳市科技计划项目(JCYJ20170307100154602;JCYJ2016031115823245)~~

摘　　要：背景:人脐带间充质干细胞具有低免疫原性,而人脐带间充质干细胞诱导分化的胰岛素分泌细胞是否保持低免疫原性尚不清楚。目的:探讨人脐带间充质干细胞向胰岛素分泌细胞诱导分化后在体外的免疫原性变化及移植到宿主后微环境对免疫原性的影响。方法:(1)采用经典改良方案诱导人脐带间充质干细胞分化为胰岛素分泌细胞,流式细胞术检测胰岛素分泌细胞免疫表型及细胞毒性实验中胰岛素分泌细胞的凋亡率;(2)CCK-8检测单向混合淋巴细胞实验中外周血单个核细胞的增殖能力;(3)小鼠腹腔及左肾包膜移植胰岛素分泌细胞后,流式细胞术及免疫组织化学检测移植部位的免疫细胞浸润情况。结果与结论:(1)人脐带间充质干细胞分化的胰岛素分泌细胞高表达HLA-ABC,低表达HLA-DR,CD40和CD80;(2)细胞毒性实验中胰岛素分泌细胞的凋亡率随预致敏脾细胞的增多而升高;(3)单向混合淋巴细胞实验中效靶比为10︰1,50︰1时胰岛素分泌细胞能抑制外周血单个核细胞增殖;(4)腹腔及左肾包膜移植胰岛素分泌细胞后移植部位淋巴细胞增多;(5)结果表明,人脐带间充质干细胞诱导分化的胰岛素分泌细胞在体外保持低免疫原性,但移植到体内后由于微环境的变化而具有一定免疫原性。BACKGROUND： Human umbilical cord mesenchymal stem cells （hUC-MSCs） have low immunogenicity and it is unclear whether insulin producing cells （IPCs） that differentiate from hUC-MSCs have low immunogenicity. OBJECTIVE：To investigate the immunogenicity of IPCs differentiating from hUC-MSCs in vitro and after IPCs transplantation into the host. METHODS： （1） The hUC-MSCs were induced to differentiate into IPCs according to the modified scheme. Flow cytometry assay was used to detect the immunophenotype and apoptotic rate of IPCs in a cytotoxicity test. （2） Cell counting kit-8 was used to detect the proliferative capacity of human peripheral blood mononuclear cells in the one-way mixed lymphocyte assay. （3） The IPCs were then transplanted into the abdominal cavity and left renal capsule of mice, and then the infiltration of immune cells was detected by flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION： The IPCs highly expressed HLA-ABC and lowly expressed HLA-DR, CD40 and CD80. The apoptosis rate of IPCs increased with the increase of pre-sensitized splenocytes in the cytotoxicity test. In the one-way mixed lymphocyte assay, IPCs inhibited the proliferation of human peripheral blood mononuclear cells when the target ratio was 10：1 and 50：1. After IPCs transplantation, the number of lymphocytes was increased in the transplanted site. In summary, our results show that IPCs that differentiate from hUC-MSCs maintain low immunogenicity in vitro,but have some immunogenicity after transplantation into the host due to microenvironment changes.

关 键 词：人脐带间充质干细胞 胰岛素分泌细胞 诱导分化 免疫原性 干细胞 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]