大量制备过继免疫治疗用γδT细胞时培养基的选择

Selection of Culture Media for the Mass Production of γδT Cells Used in Adoptive Immunotherapy

目的选择适于过继免疫治疗用γδT细胞大量制备的培养基。方法采用不同培养基(RPMI-1640、AIM-V和OpTmizer,分别添加或者不添加自体血清)对γδT细胞进行培养,比较细胞存活率、纯度、扩增效率和生物学功能。结果仅未添加血清的RPMI-1640培养基扩增的细胞存活率随培养时间增加略有下降。不论添加血清与否,AIM-V和OpTmizer培养基扩增的细胞纯度都始终高于RPMI-1640。第2周时,未添加血清的OpTmizer培养基扩增的细胞纯度和扩增效率都显著高于RPMI-1640培养基和AIM-V培养基(P均<0.05)。不同培养基扩增细胞表达的CD107a和分泌的肿瘤坏死因子-α差异均没有统计学意义(P均>0.05),但RPMI-1640培养的细胞对Daudi细胞的杀伤效率显著低于OpT-mizer和AIM-V培养的细胞(P均<0.05)。结论 OpTmizer培养基因其低血清依赖性、高扩增效率和扩增细胞良好的生物学功能,更适用于临床过继免疫治疗用γδT细胞的大量培养。

Objective To select the optimal culture media for the mass production of γδT cells used in adoptive immunotherapy.Methods Three different culture media（RPMI-1640,AIM-V,and OpTmizer,with or without autologous serum） were used to culture γδT cells.The survival rate,purity,proliferation efficiency,and biological functions of the expanded γδT cells were examined and compared.Results The survival rate of γδT cells expanded in RPMI-1640 decreased over culture time.The purities of γδT cells cultured in AIM-V or OpTmizer with or without serum were higher than those cultured in RPMI-1640.After two weeks of culture in the absence of serum,the purity and proliferation efficiency of γδT cells cultured in OpTmizer were significantly higher than those cultured in RPMI-1640（P0.05） and AIM-V（P0.05）.γδT cells in different culture media had similar CD107a expression and tumor necrosis factor-α production（P0.05）.However,cells expanded in RPMI-1640 exhibited significantly weaker cytotoxicity against Daudi lymphoma cells than those expanded in OpTmizer（P0.05） and AIM-V（P0.05）.Conclusion Due to low serum-dependence,high proliferation efficiency,and favorable biology function of expanded cells,OpTmizer is the most suitable medium for the mass production of γδT cells used in adoptive immunotherapy.