LymphoONE T-Cell Expansion Xeno-Free Medium is specifically formulated for expansion and transduction of human T lymphocytes. It has been optimized to enable efficient T-cell proliferation with or without the addition of human serum or plasma. LymphoONE medium contains no protein other than pharmaceutical-grade human serum albumin and recombinant human insulin, and provides superior performance relative to other commonly used xeno-free media.LymphoONE T-Cell Expansion Xeno-Free Medium is specifically formulated for expansion and transduction of human T lymphocytes. It 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 xeno-free media.Conventional culture systems typically require the addition of serum (or plasma) in order to achieve sufficient quantities of T cells for clinical research applications, but the availability of autologous serum varies, and it has been demonstrated that T cells expanded in the absence of serum can offer improved therapeutic performance (Janetzki et al. 2010; Medvec et al. 2018). By enabling robust expansion in the absence of human serum or plasma, LymphoONE medium is well-suited for immunotherapy research involving T cells.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.As demonstrated by the data below, expansion in LymphoONE medium also yields higher proportions of naïve T cells as compared to other commonly used media, with or without the addition of serum.Phenotypic analysis of T cells expanded using various culture media in the presence or absence of 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. Expansion in LymphoONE medium resulted in higher proportions of cells expressing markers associated with a naïve T-cell phenotype (CD45RA+/CCR7+), with or without addition of serum. More Less