羊膜间充质干细胞的免疫学特性研究

发布时间：2018-01-04 15:06

本文关键词：羊膜间充质干细胞的免疫学特性研究　出处：《昆明医学院》2009年硕士论文　论文类型：学位论文

【摘要】： 目的:从羊膜中分离、培养间充质干细胞(MSCs),分析其免疫原性及对淋巴细胞转化的影响,以期为寻找更易获得且易于培养和纯化的MSCs新来源及临床提高造血干细胞移植(HSCT)成功率、减少移植物抗宿主病(GVHD)的发生提供实验依据。方法:采用组织块培养法从羊膜中分离、培养MSCs;用流式细胞术分析羊膜MSCs表面的HLA-A,B,C、HLA-DR以及T淋巴细胞活化所需共刺激分子CD80、CD86、CD40的表达;用混合淋巴细胞反应检测羊膜MSCs刺激同种异体T淋巴细胞增殖的能力;应用CCK-8法观察羊膜MSCs对成人外周血淋巴细胞转化的影响。结果:羊膜MSCs具有低免疫原性,其细胞表面只表达低水平的HLA-A,B,C,不表达HLA-DR以及CD80、CD86、CD40,不会引起同种异体T淋巴细胞的增殖反应,能够抑制成人外周血淋巴细胞的转化,且这种抑制作用具有剂量依赖性。当羊膜MSCs与淋巴细胞的比例为1:10和1:50时均未能刺激同种异体T淋巴细胞增殖。当羊膜MSCs与淋巴细胞的比例为1:1时对淋巴细胞转化的平均抑制率为88.92%;当羊膜MSCs与淋巴细胞的比例为1:2时对淋巴细胞转化的平均抑制率为83.38%:当羊膜MSCs与淋巴细胞的比例为1:4时对淋巴细胞转化的平均抑制率为62.10%;当羊膜MSCs与淋巴细胞的比例为1:8时对淋巴细胞转化的平均抑制率为32.22%;当羊膜MSCs与淋巴细胞的比例为1:16时对淋巴细胞转化的平均抑制率为17.35%。结论:从羊膜中可以成功分离、培养出MSCs,利用其低免疫原性及对淋巴细胞的免疫调节作用,有望在诱导免疫耐受、减少和预防GVHD中发挥重要作用,提高HSCT的成功率,减少GVHD的发生。
[Abstract]:Objective: to isolate from the amniotic membrane in cultured mesenchymal stem cells (MSCs), analysis of the impact of its immunogenicity and on lymphocyte transformation, in order to find a more accessible and easy to MSCs new source and clinical training and purification of improving hematopoietic stem cell transplantation (HSCT) success rate, reduce graft-versus-host disease (GVHD) to provide experimental basis.
Methods: using tissue separation, from amnion cultured MSCs; flow cytometry analysis of amniotic membrane surface of MSCs HLA-A, B, C, HLA-DR and T lymphocyte activation required costimulatory molecules CD80, CD86, CD40 expression ability; Needling proliferation of allogeneic T lymphocytes induced by MSCs detection of amniotic membrane mixed lymphocyte reaction the effects of MSCs on amniotic membrane; adult peripheral blood lymphocyte transformation by CCK-8 method.
Results: MSCs amniotic membrane has low immunogenicity, the cell surface expressed only low levels of HLA-A, B, C, HLA-DR and CD80 expression, CD86, CD40, will not cause the proliferation of allogeneic T lymphocytes, can inhibit the conversion of adult peripheral blood lymphocytes, and this inhibition is dose dependent. When the amniotic membrane MSCs and lymphocyte ratio of 1:10 and 1:50 were unable to stimulate allogeneic T lymphocyte proliferation. When the amniotic membrane MSCs and lymphocyte ratio average inhibition 1:1 on lymphocyte transformation rate was 88.92%; when the amniotic membrane MSCs and lymph cell ratio was 1:2 on average inhibition of lymphocyte transformation rate was 83.38% when: MSCs and lymphocyte ratio of amniotic membrane average inhibition 1:4 on lymphocyte transformation rate was 62.10%; when the amniotic membrane MSCs and lymphocyte ratio was 1:8 on average inhibition of lymphocyte transformation rate was 32.22%; When the proportion of MSCs and lymphocyte in amniotic membrane was 1:16, the average inhibition rate of lymphocyte transformation was 17.35%.
Conclusion: MSCs can be successfully isolated from amniotic membrane and cultured, and its low immunogenicity and immunomodulatory effect on lymphocytes are expected to play an important role in inducing immune tolerance, reducing and preventing GVHD, improving the success rate of HSCT and reducing the occurrence of GVHD.

【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

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