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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。
Objective To study the isolation and culture of human amniotic membrane-derived mesenchymal stem cells and their immunoregulatory effects. Methods Mesenchymal stem cells were isolated and cultured from amniotic membrane, and the surface markers were detected by morphological structure and flow cytometry to identify their purity. To establish a two-way mixed lymphocyte reaction system, mitomycin-treated amnion cells were added according to different effective target ratios (hAMSCs / PBMC), and the lymphocyte proliferation rate was measured by 3H incorporation method. The effects of mesenchymal stem cells Effect of cell proliferation. Results Mesenchymal stem cells (MSCs) were successfully cultured from amniotic membrane and the cells grew adherently. They could proliferate and pass on in vitro in a short period. Cells with similar surface markers to BMSCs co-cultured with mixed lymphocyte reaction inhibited Lymphocyte proliferation, the proliferation inhibition rate is proportional to the number of cells added. Conclusion Amniotic mesenchymal stem cells can inhibit the proliferation of lymphocytes in the mixed lymphocyte reaction system and have a negative regulation on the allogeneic immune response.