胎盘间充质干细胞与免疫抑制药物体外联合应用对单向淋巴细胞混合反应的作用

Effect of combined application of placenta-derived mesenchymal stem cells and immunosuppressive drug in vitro on one-way mixed lymphocyte reaction

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 the placenta-derived mesenchymal stem cells（ pMSCs）, and to detect immunomodulatory effects of pMSCs or pMSCs combinated with immunosuppressive drugs（ cyclospo- fine A/mycophenolic acid）on one-way mixed lymphocyte reaction. Methods Tissue culture method was used to extract pMSCs. Cell surface markers was detected by flow cytometry, and type II collagen immuno- histochemical staining and toluidine blue staining were used to detect the ability of pMSCs to differentiate in- to chondrocytes. After pMSCs or pMSCs combinated with cyclosporine A/mycophenolic acid were added in- to one-way lymphocyte reaction, the proliferation of T cells was detected by MTT. Results pMSCs were suc- cessfully extracted from placenta. They positively expressed CD29 and CD44. Type II collagen staining and toluidine blue staining presented positive results. Application of pMSCs alone could inhibit the proliferation of T cells in one-way mixed lymphocyte reaction. But once it was combined with cyclosporine A, the inhibi- tion efficiency was decreased. On the contrary, the inhibition efficiency was increased when it was combined with mycophenolic acid. Conclusions pMSCs can be successfully extracted from placenta by tissue culture method. They have a negative immunomodulatory effect on one-way mixed lymphocyte reaction. When pM- SCs was combined with different immune suppressive drugs, the resultant immunomodulatory effects were different.