摘要目的:研究外加中等强度静磁场是否可以辅助提高人T细胞体外扩增效率。方法:采集健康志愿者外周血，利用淋巴细胞分离液及密度梯度离心法分离人外周血单核细胞（PBMCs），利用MACS磁珠分选法阴性分选人CD3 + T细胞，加入人T细胞活化珠及重组人白介素IL2体外刺激培养T细胞1周。同等培养条件下，细胞培养板置入钕铁硼产生的静磁场中（场强0.22±0.01T），比较两种环境下T细胞体外扩增效率，流式细胞术检测人T细胞活化及耗竭标志物表达，体外扩增后的T细胞与人肝癌细胞Hep3B细胞共培养，比较T细胞抑制人肝癌细胞增殖的能力。 结果:相比常规培养，外加静磁场可显著提高人T细胞体外活化程度及增殖倍数，使其1周的增殖倍数由6.11倍提高到14.83倍（ P<0.05），且T细胞表达更多的活化标志物CD25，CD137及细胞增殖标志物Ki-67，T细胞耗竭标志物PD-1表达量显著降低（ P<0.05）。T细胞抑制Hep3B细胞增殖实验结果显示，当T细胞与肝癌细胞按1∶10共培养时，外加静磁场下扩增的T细胞平均抑制率为30.1%，而无磁场环境下扩增的T细胞其抑制率为12.5%， P<0.05。 结论:外加中等强度静磁场可以提高人T细胞体外扩增效率，加速T细胞的活化与增殖，减缓其耗竭程度并增强其抑制人肝癌细胞增殖的能力。

abstractsObjective:To study whether moderate-intensity static magnetic field can help improve the expansion efficiency of human T lymphocytes in vitro.Methods:Peripheral blood samples were collected from healthy volunteers and used for separation of human peripheral blood mononuclear cells (PBMCs) using lymphocyte separation medium and density gradient centrifugation. Human CD3 + T lymphocytes were isolated by negative selection using magnetically activated cell sorting (MACS) . In-vitro culture of the T cells was performed with addition of human T cell activation beads and recombinant human interleukin IL2 stimulation for one week. Under the same culture conditions, the cell culture plate was placed in a static magnetic field generated by NdFeB magnets (field strength, 0.22±0.01T) . The in-vitro expansion efficiency of T cells was compared between these two settings. Flow cytometry was used to detect the markers of human T cell activation and depletion. The T cells expanded in-vitro were co-cultured with human hepatocarcinoma Hep3B cells. The inhibitory effect against human hepatocarcinoma cells was compared between T cells cultured under the two conditions. Results:Compared with conventional culture, external static magnetic field significantly increased the human T cells activation and fold of proliferation in vitro, leading to a one-week fold increase from 6.11 to 14.83 fold ( P<0.05) . Higher expression of T cell activation markers (CD25 and CD137) and cell proliferation marker Ki-67, and lower expression of T cell exhaustion marker PD-1 ( P<0.05) . The experiment of T cell inhibition against Hep3B cell proliferation showed that when the T cells and hepatocarcinoma cells were co-cultured at a ratio of 1: 10, the mean inhibition rate was 30.1% for T cells expanded under an external static magnetic field, compared with those expanded without a magnetic field environment (12.5%, P<0.05) . Conclusion:Moderate-intensity static magnetic field can improve the expansion efficiency of human T lymphocytes in vitro, accelerate the T cell activation and proliferation, retard the T cell exhaustion, and enhance their inhibitory effect against proliferation of human hepatocarcinoma cells.