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不同培养基环境对人T细胞体外培养的影响。

Influence of various medium environment to in vitro human T cell culture.

作者信息

Xu Hao, Wang Na, Cao Wenyue, Huang Liang, Zhou Jianfeng, Sheng Lingshuang

机构信息

Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, Hubei, China.

出版信息

In Vitro Cell Dev Biol Anim. 2018 Sep;54(8):559-566. doi: 10.1007/s11626-018-0273-3. Epub 2018 Jul 12.

Abstract

Nowadays, adoptive T cell immunotherapy is emerging as a novel and potent treatment for cancer. To prepare enough effective T cells for treatment use, their rapid expansion is favorable. Our study compared 6 commonly used cultural media for human T cells, including serum-containing media and serum-free media, namely RPMI 1640, IMDM, Gibco OpTmizer CTS T Cell Expansion SFM, Gibco AIM-V Medium CTS, LONZA X-VIVO 15, and StemSpan SFEM with or without Dynabeads Human T-Activator CD3/CD28, on in vitro T cell expansion, apoptosis, and immune phenotype. Our study results suggest that serum-free media provide better proliferation environment for T cells. Among the 3 serum-free media, we identify OpTmizer and AIM-V as better T cell culture environments compared with X-VIVO as T cells are proved to have higher viability in the first two media. Besides, we found that in vitro human T cells keep relatively resting status among non-CD3/CD28 groups, since they have weak proliferation and apoptosis abilities. The phenotypes of T cells in different cultural environments over time indicate T cells maturation during culture duration. These results provide a firm foundation of adoptive T cell immunotherapy.

摘要

如今,过继性T细胞免疫疗法正成为一种新型且有效的癌症治疗方法。为制备足够数量的有效T细胞用于治疗,T细胞的快速扩增是有利的。我们的研究比较了6种常用于人T细胞的培养基,包括含血清培养基和无血清培养基,即RPMI 1640、IMDM、Gibco OpTmizer CTS T细胞扩增无血清培养基、Gibco AIM-V培养基CTS、LONZA X-VIVO 15以及添加或不添加Dynabeads人T细胞激活剂CD3/CD28的StemSpan SFEM,观察其对体外T细胞扩增、凋亡及免疫表型的影响。我们的研究结果表明,无血清培养基为T细胞提供了更好的增殖环境。在3种无血清培养基中,我们发现与X-VIVO相比,OpTmizer和AIM-V作为更好的T细胞培养环境,因为T细胞在前两种培养基中的活力更高。此外,我们发现体外人T细胞在非CD3/CD28组中保持相对静止状态,因为它们的增殖和凋亡能力较弱。不同培养环境下T细胞表型随时间的变化表明T细胞在培养过程中逐渐成熟。这些结果为过继性T细胞免疫疗法奠定了坚实基础。

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