人羊膜间充质干细胞抑制异体淋巴细胞增殖的实验研究

目的研究人羊膜间充质干细胞分离培养及免疫调节作用。方法从羊膜中分离和培养间充质干细胞,并通过形态学的结构特征及流式细胞术检测其表面标志以鉴定其纯度。建立双向混合淋巴细胞反应体系,根据不同效靶比E/T（hAMSCs/PBMC）加入丝裂霉素处理过的羊膜细胞,用3H掺入法测定淋巴细胞增殖率,观察间充质干细胞对淋巴细胞增殖的影响。结果从羊膜组织成功培养出间充质干细胞,细胞贴壁生长,在体外短期内可以稳定增殖和传代,细胞具有与骨髓间充质干细胞相似的表面标志,与混合淋巴细胞反应体系共同培养抑制了淋巴细胞增殖,其增殖抑制率与加入的细胞数量成正比。结论羊膜间充质干细胞可以抑制混合淋巴细胞反应体系中淋巴细胞增殖,对同种异体免疫反应具有负调节作用。

Suppressive Effect of Human Amniotic Mesenchymal Stromal Cells on Allogeneic Lymphocytes Proliferation

Objective To study the fractional cultivation method and the immunomodulatory functions of human amniotic mesenchymal stromal cells （hAMSCs） in vitro. Methods Amniotic mesenchymal stromal cells from human were cultured with tissue piece method, passaged by trypsin digestion and identified with immunocytochemistry and fluorescence-activated cell sorter （FACS）. HAMSCs treated by mitomyein according to different effective/target ratios（hAMSCs/peripheral blood monouclear cells） were added in the mixed lymphocyte reaction（MLR） system. To determine the growth rate of lymphocyte with 3H-TdR and observe the effect of hAMSCs on lymphocyte multiplication. Results HAMSCs migrated from explants and primary culture were established; they could multiply and expand steadily in a short time, and they could be passaged by trypsin digestion. The surface marker of hAMSCs was the same as that of bone marrow mesenchymal stem cells, hAMSCs could inhibit lymphocyte proliferation in MLR system and the influence on proliferation of lymphocyte related to the ratio of hAMSCs and lymphocyte. Conclusion HAMSCs can inhibit lymphocyte proliferation in MLR system and have down regulation effect in alloimmunity-reaction.