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.
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References
Bruserud O, Tronstad KJ (2005) In vitro culture of human osteosarcoma cell lines: a comparison of functional characteristics for cell lines cultured in medium without and with fetal calf serum. J Cancer Res Clin Oncol 131:377–384. https://doi.org/10.1007/s00432-004-0650-z
Chang ZL, Silver PA (2015) Identification and selective expansion of functionally superior T cells expressing chimeric antigen receptors. J Transl Med 13:161. https://doi.org/10.1186/s12967-015-0519-8
Dai H, Wang Y, Lu X (2016) Chimeric antigen receptors modified T-cells for cancer therapy. J Natl Cancer Inst 108. https://doi.org/10.1093/jnci/djv439
Fesnak AD, June CH (2016) Engineered T cells: the promise and challenges of cancer immunotherapy. Nat Rev Cancer 16:566–581. https://doi.org/10.1038/nrc.2016.97
Golubovskaya V (2016) Different subsets of T cells, memory, effector functions, and CAR-T immunotherapy. Cancers (Basel) 8. https://doi.org/10.3390/cancers8030036
Jameson SC (2009) Diversity in T cell memory: an embarrassment of riches. Immunity 31:859–871. https://doi.org/10.1016/j.immuni.2009.11.007
Janetzki S, Price L, Britten CM et al (2010) Performance of serum-supplemented and serum-free media in IFNgamma Elispot assays for human T cells. Cancer Immunol Immunother 59:609–618. https://doi.org/10.1007/s00262-009-0788-2
Jeon MK, Lim JB (2010) Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design. BMC Biotechnol 10:70. https://doi.org/10.1186/1472-6750-10-70
Klaver Y, van Steenbergen SC, Sleijfer S et al (2016) T cell maturation stage prior to and during GMP processing informs on CAR T cell expansion in patients. Front Immunol 7:648. https://doi.org/10.3389/fimmu.2016.00648
Kochenderfer JN, Feldman SA, Zhao Y et al (2009) Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. J Immunother 32:689–702. https://doi.org/10.1097/CJI.0b013e3181ac6138
Lu TL, Pugach O, Somerville R et al (2016) A rapid cell expansion process for production of engineered autologous CAR-T cell therapies. Hum Gene Ther Methods 27:209–218. https://doi.org/10.1089/hgtb.2016.120
Lutz MB (2007) Factors influencing the generation of murine dendritic cells from bone marrow: the special role of fetal calf serum. Immunobiology 212:855–862. https://doi.org/10.1016/j.imbio.2007.09.001
Mailer RKW, Gisterå A, Polyzos KA, Ketelhuth DFJ, Hansson GK (2017) Hypercholesterolemia enhances T cell receptor signaling and increases the regulatory T cell population. Sci Rep 7:15655. https://doi.org/10.1038/s41598-017-15546-8
Moretta A, Ciccone E, Pantaleo G et al (1989) Surface molecules involved in the activation and regulation of T or natural killer lymphocytes in humans. Immunol Rev 111:145–175
Sato K, Kondo M, Sakuta K et al (2009) Impact of culture medium on the expansion of T cells for immunotherapy. Cytotherapy 11:936–946. https://doi.org/10.3109/14653240903219114
Schiller CB, Braciak TA, Fenn NC et al (2016) CD19-specific triplebody SPM-1 engages NK and γδ T cells for rapid and efficient lysis of malignant B-lymphoid cells. Oncotarget 7:83392–83408. https://doi.org/10.18632/oncotarget.13110
Sommermeyer D, Hudecek M, Kosasih PL et al (2016) Chimeric antigen receptor-modified T cells derived from defined CD8+ and CD4+ subsets confer superior antitumor reactivity in vivo. Leukemia 30:492–500. https://doi.org/10.1038/leu.2015.247
Sutton KS, Dasgupta A, McCarty D et al (2016) Bioengineering and serum free expansion of blood-derived γδ T cells. Cytotherapy 18:881–892. https://doi.org/10.1016/j.jcyt.2016.04.001
Turtle CJ, Hanafi LA, Berger C et al (2016) CD19 CAR-T cells of defined CD4+:CD8+ composition in adult B cell ALL patients. J Clin Invest 126:2123–2138. https://doi.org/10.1172/JCI85309
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Xu, H., Wang, N., Cao, W. et al. Influence of various medium environment to in vitro human T cell culture. In Vitro Cell.Dev.Biol.-Animal 54, 559–566 (2018). https://doi.org/10.1007/s11626-018-0273-3
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DOI: https://doi.org/10.1007/s11626-018-0273-3