LymphoONE T-Cell Expansion Xeno-Free Medium

Brand	Takara Bio

LymphoONE T-Cell Expansion Xeno-Free Medium is specifically formulated for expansion and transduction of human T lymphocytes. The medium has been optimized to enable efficient T-cell proliferation with or without the addition of human serum or plasma. LymphoONE medium contains no proteins other than pharmaceutical-grade human serum albumin and recombinant human insulin, and provides superior performance relative to other commonly used serum-free media. T-cell activation using LymphoONE T-Cell Expansion Xeno-Free Medium in combination with anti-CD3 antibody, IL-2, and RetroNectin reagent results in enhanced T-cell expansion and yields higher proportions of naïve T cells relative to approaches employing IL-2 in combination with anti-CD3 and anti-CD28 antibodies. LymphoONE medium also contains L-glutamine and streptomycin, and is confirmed to have low endotoxin levels (<0.3 EU/ml).

Overview

Xeno-free medium that enables efficient T-cell expansion with or without the addition of serum or plasma
Supplemented with human serum albumin, recombinant human insulin, L-glutamine, and streptomycin
Endotoxin tested (endotoxin <0.3 EU/ml)
Compatible with CultiLife culture bags for large-scale expansion of T cells
Anti-CD3/IL-2 T-cell activation in the presence of RetroNectin reagent* enhances proliferation and leads to a higher proportion of naïve T cells

*T-cell expansion by RetroNectin stimulation for uses other than research requires a license from Takara Bio Inc.

Applications

Expansion of T cells
Gene transfer by lentiviral or retroviral transduction

LymphoOne medium enables efficient T-cell expansion without the addition of serum or plasma

T-cell expansion comparison using various serum-free culture media. Peripheral blood mononuclear cells (PBMCs) were isolated from blood obtained from two healthy donors and cultured for four days under stimulation by anti-CD3 monoclonal antibody and IL-2 in CultiLife 215 Culture Bags using LymphoONE T-Cell Expansion Xeno-Free Medium, GT-T551 (Takara Bio), X-VIVO 15 (Lonza), or AIM V (Thermo Fisher Scientific). The cells were then cultured at appropriate dilutions for an additional 10 days in each of the respective media supplemented with IL-2. By Day 14, cells cultured in LymphoONE medium exhibited higher fold expansion than cells cultured in either of the other three media.

Supplementation of LymphoONE medium with human serum results in greater T-cell expansion and yields higher proportions of naive T cells

T-cell expansion via stimulation by anti-CD3 antibody, IL-2, and RetroNectin in the presence or absence of human serum. Peripheral blood mononuclear cells (PBMCs) were cultured for four days under stimulation by anti-CD3 monoclonal antibody, IL-2, and RetroNectin reagent in CultiLife 215 Culture Bags with or without addition of human serum using LymphoONE T-Cell Expansion Xeno-Free Medium, X-VIVO 15 (Lonza), or AIM V (Thermo Fisher Scientific). The cells were then cultured at appropriate dilutions with or without serum for an additional six days in each of the respective media supplemented with IL-2. Human serum was added at a concentration of 1 from Day 0 to Day 7, and then at a concentration of 0.5 from Day 7 to Day 10. Top panel. By Day 10, cells cultured in LymphoONE medium exhibited higher fold expansion than cells cultured in either of the other two media, regardless of whether serum was added. Bottom panel. Expansion in LymphoONE medium also resulted in higher proportions of cells expressing markers associated with naïve T-cell phenotype (CD45RA+/CCR7+).

Serum-free T-cell expansion protocol schematic

T-cell expansion protocol using RetroNectin reagent, LymphoOne xeno-free medium, CultiLife bags, and anti-CD3 mAb.

Reference

Janetzki, S. et al. Performance of serum-supplemented and serum-free media in IFNgamma Elispot Assays for human T cells. Cancer Immunol. Immunother. 59, 609–18 (2010).

Medvec, A. R. et al. Improved expansion and in vivo function of patient T cells by a serum-free medium. Mol. Ther. Methods Clin. Dev. 8, 65–74 (2018).

LymphoONE T-Cell Expansion Xeno-Free Medium

Additional information