目的 通过共培养异源的人骨髓间充质干细胞和树突状细胞活化的细胞因子激活的杀伤细胞(DC-CIK细胞),观察DC-CIK细胞的细胞周期,初步探讨骨髓间充质干细胞的免疫调节机制.方法 采用Ficoll密度梯度离心法分离、培养健康骨髓间充质干细胞和外周血单个核细胞,用多种细胞因子诱导培养外周血单个核细胞中贴壁和悬浮细胞分别为树突状细胞和杀伤细胞.将骨髓间充质干细胞与DC-CIK细胞以1∶5,1∶10,1∶20共培养4 d,采用流式细胞术检测DC-CIK细胞的细胞周期.结果 骨髓间充质干细胞呈长梭形,表达CD+29CD+444双阳性的细胞为93.6%,实验选用第三代后的细胞.在培养第9天树突状细胞表达CD1α人类白细胞Ⅱ类抗原(HLA-DR+)]双阳性细胞数为98.5%.DC-CIK细胞表达CD+3CD+56的细胞数为(29.2±12.2)%.骨髓间充质干细胞使DC-CIK细胞滞留在G0/G1期,且与细胞比例呈正相关.以1∶5,1∶10,1∶20的比例共培养4 d的DC-CIK细胞的G0/G1期和S期分别为(91.0±3.3)%和(2.8±0.6)%,(88.7±3.0)%和(5.1±2.8)%,(83.7±1.7)%和(8.7±3.4)%.对照组(同步化DC-CIK细胞)为(73.4±1.0)%和(17.4±0.6)%.结论 骨髓间充质干细胞可通过抑制DC-CIK细胞的细胞周期而发挥免疫调节作用.%Objective To explore the effect of allogeneic human bone marrow derived mesenchymal stem cells (hMSCs) on cell cycle of DC-CIK cells (dendritic cell activated cytokine-induced killer cells),and the mechanism of immunoregulation induced by hMSCs. Methods The hMSCs and peripheral blood mononuclear cells were separated by Ficoll gradient centrifugation. The DC and CIK cells from peripheral blood were induced by multi-cytokine. The hMSCs were co-cultured with synchronously DC-CIK cells accordsults The hMSCs displayed a fibroblast-like morphology, in which the double positive cells of CD+29 and CD+44 were 93.6% after 3 passages. The expression of CD1α + HLA-DR+ in DC were 98.5% at 9 days. The CD3+ CD+56 double positive cells in DC-CIK cells were (29.2 ± 12.2)%. The DC-CIK cells in the presence 2.8)% ;(83.7± 1.7)% and (8.7± 3.4)%, respectively. In control group(DC-CIK cells), it is (73.4±1.0)% and(17.4±0.6)%. Conclusion hMSCs can exert an influence on immunoregulation by inhibiting cell cycles of DC-CIK cells.
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