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[用于过继性免疫治疗的γδT细胞大规模生产的培养基选择]

[Selection of culture media for the mass production of gamma delta T cells used in adoptive immunotherapy].

作者信息

Zhou Jian-Hua, Kang Ning, Cui Lian-Xian, He Wei

机构信息

Department of Immunology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2011 Dec;33(6):644-8.

Abstract

OBJECTIVE

To select the optimal culture media for the mass production of gamma delta T 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 gamma delta T cells. The survival rate, purity, proliferation efficiency, and biological functions of the expanded gamma delta T cells were examined and compared.

RESULTS

The survival rate of gamma delta T cells expanded in RPMI-1640 decreased over culture time. The purities of gamma delta T 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 gamma delta T cells cultured in OpTmizer were significantly higher than those cultured in RPMI-1640 (P < 0.05) and AIM-V (P < 0.05). gamma delta T cells in different culture media had similar CD107a expression and tumor necrosis factor-alpha production (P > 0.05). However, cells expanded in RPMI-1640 exhibited significantly weaker cytotoxicity against Daudi lymphoma cells than those expanded in OpTmizer (P < 0.05) and AIM-V (P < 0.05).

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 gamma delta T cells used in adoptive immunotherapy.

摘要

目的

筛选用于过继性免疫治疗中大量生产γδT细胞的最佳培养基。

方法

使用三种不同的培养基(RPMI-1640、AIM-V和OpTmizer,添加或不添加自体血清)培养γδT细胞。检测并比较扩增后的γδT细胞的存活率、纯度、增殖效率和生物学功能。

结果

在RPMI-1640中扩增的γδT细胞的存活率随培养时间下降。在添加或不添加血清的AIM-V或OpTmizer中培养的γδT细胞的纯度高于在RPMI-1640中培养的细胞。在无血清条件下培养两周后,在OpTmizer中培养的γδT细胞的纯度和增殖效率显著高于在RPMI-1640(P<0.05)和AIM-V(P<0.05)中培养的细胞。不同培养基中的γδT细胞具有相似的CD107a表达和肿瘤坏死因子-α产生(P>0.05)。然而,在RPMI-1640中扩增的细胞对Daudi淋巴瘤细胞的细胞毒性明显弱于在OpTmizer(P<0.05)和AIM-V(P<0.05)中扩增的细胞。

结论

由于OpTmizer血清依赖性低、扩增细胞增殖效率高且生物学功能良好,是用于过继性免疫治疗中大量生产γδT细胞的最合适培养基。

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