The state of T cells before cryopreservation: Effects on post-thaw proliferation and function
The state of T cells before cryopreservation: Effects on post-thaw proliferation and function
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DOI：
10.1016/j.cryobiol.2017.08.008
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发表时间：
2017-12-01
期刊：
影响因子：
2.7
通讯作者：
Gao, Yingtang
中科院分区：
文献类型：
--
作者：
Luo, Ying;Wang, Peng;Gao, Yingtang
目的：我们的目的是评估冻存前T细胞的状态对解冻后的增殖能力，表型和功能respons.Methods：外周血单个核细胞（PBMC）从肝细胞癌（HCC）患者分离的效果，和T细胞冷冻在细胞培养过程中根据我们的实验设计。结果：各组冻存细胞解冻后存活率均>90%，冻存后细胞增殖能力、T细胞表面标志物表达、IFN-γ和IL-10分泌均高于对照组。在细胞培养过程中第4、8或12天冷冻保存的淋巴细胞允许恢复24小时，而新鲜分离时冷冻保存的淋巴细胞允许恢复72小时。再培养期后，在T细胞培养期间第4、8或12天的冷冻保存未发现改变T细胞亚群。T细胞培养期间第12天细胞冻存时，NMT和Treg细胞的比例没有变化。结论：冻存前T细胞的状态对T细胞解冻后的增殖能力、表型及IFN-γ、IL-10的分泌均有影响。在细胞培养的第8或12天冷冻保存淋巴细胞可能是T细胞免疫治疗的最佳选择。(C)2017爱思唯尔公司All rights reserved.
Aim: We aim to assess the effect of the state of T cells before cryopreservation on the post-thaw proliferative capacity, phenotype and functional response.Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from a hepatocellular carcinoma (HCC) patient, and the T cells were frozen during cell culture according to our experimental design. After a period of re-culture, the proliferative capacity of the cryopreserved cells, the expression of T cell surface markers and the secretion of IFN-gamma and IL-10 were assayed.Results: There was >90% cell viability after thaw in every group. Lymphocytes cryopreserved at day 4, 8 or 12 during the cell culture were allowed to recover for 24 h, whereas lymphocytes cryopreserved while freshly isolated were allowed to recover for 72 h. After the period of re-culture, cryopreservation at day 4, 8 or 12 during T cell culture was not found to alter the T cell subpopulation. The proportions of NMT and Treg cells were unchanged when cells were cryopreserved at day 12 during T cell culture. IFN-gamma secretion was not impacted by cryopreservation, and IL-10 secretion was significantly decreased when cells were cryopreserved at day 8 or 12 during T cell culture.Conclusion: The state of T cells before cryopreservation has effects on the post-thaw proliferation capacity, the phenotype and the secretion of IFN-gamma and IL-10. Cryopreservation of lymphocytes at day 8 or 12 during the cell culture may be the best choice for T cell immunotherapy. (C) 2017 Elsevier Inc. All rights reserved.