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。
Objective 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.
作者
雷虹
唐倩倩
李艳
马锋
王浩华
刘仕琪
吴荣谦
吕毅
Lei Hong;Tang Qianqian;Li Yan;Ma Feng;Wang Haohua;Liu Shiqi;Wu Rongqian;Lyu Yi(Affiliated Children’s Hospital of Xi’an Jiaotong University,Shaanxi Institute of Pediatric Diseases,Shaanxi Provincial Key Laboratory of Precision Medicine for Pediatric Diseases,Xi’an 710003,China;National and Local Joint Engineering Research Center for Precision Surgery and Regenerative Medicine,First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China;Department of Neonatal Surgery,Affiliated Children’s Hospital of Xi’an Jiaotong University,Xi’an 710003,China)
出处
《中华生物医学工程杂志》
CAS
2022年第4期381-387,共7页
Chinese Journal of Biomedical Engineering
基金
国家自然科学基金重大研究计划(92048202)
国家自然科学基金(82170676)
西安交通大学基本科研业务费(xjh012020018)。
关键词
T淋巴细胞
磁场
体外扩增
T细胞治疗
T lymphocytes
Magnetic field
In-vitro expansion
T cell therapy
