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目的提取并鉴定胎盘间充质干细胞,检测胎盘间充质干细胞对单向淋巴细胞混合反应的免疫调节作用,并将胎盘间充质干细胞与免疫抑制药物(环孢素A或霉酚酸)联合应用,检测二者联合后对单向淋巴细胞混合反应的作用。方法组织块培养法提取胎盘间充质干细胞,流式细胞仪检测细胞表面标志,II型胶原免疫组化染色及甲苯氨蓝染色检测胎盘间充质干细胞向软骨细胞分化的能力。MTT检测单独应用胎盘间充质干细胞或与环孢素A或霉酚酸联合应用后,单向淋巴细胞反应中T细胞的增殖情况。结果成功提取胎盘间充质干细胞,其表面阳性表达CD29和CD44。II型胶原及甲苯氨蓝染色均呈阳性。单独应用胎盘间充质干细胞可抑制单向淋巴细胞混合反应中T细胞的增殖。与环孢素A联合后该作用减弱,与霉酚酸联合后该作用增强。结论组织块培养法可成功提取胎盘间充质干细胞。该细胞对单向淋巴细胞混合反应有负向免疫调节作用。胎盘间充质干细胞与不同的免疫抑制药物联合对单向淋巴细胞混合反应产生的作用是有差异的。
OBJECTIVE: To extract and identify placental mesenchymal stem cells (MSCs) to detect the immunomodulatory effect of placental mesenchymal stem cells (MSCs) on unidirectional lymphocyte mixed response and to combine placental mesenchymal stem cells with immunosuppressive drugs (cyclosporin A or mycophenolic acid) Application, testing the combination of the two unidirectional lymphocyte reaction. Methods Tissue culture method was used to extract placental mesenchymal stem cells. Flow cytometry was used to detect cell surface markers. Type II collagen immunohistochemistry and toluidine blue staining were used to detect the differentiation of placental mesenchymal stem cells into chondrocytes. MTT assay of placental mesenchymal stem cells alone or in combination with cyclosporine A or mycophenolic acid in unidirectional lymphocyte reaction of T cell proliferation. Results Successful extraction of placental mesenchymal stem cells, the positive expression of CD29 and CD44 on the surface. Type II collagen and toluidine blue staining were positive. Placental mesenchymal stem cells alone can inhibit T cell proliferation in unidirectional lymphocyte mixed reactions. This effect was weakened after combined with cyclosporine A, which increased with mycophenolic acid. Conclusion Tissue culture can successfully extract placental mesenchymal stem cells. The cells have a negative immunomodulatory effect on unidirectional lymphocyte mixed responses. The role of placental mesenchymal stem cells in combination with different immunosuppressive drugs on unidirectional lymphocyte mixed responses is different.