Figure 1. ImmunoCult™-XF Supports Equivalent or Faster T Cell Expansion Compared to Commercial Alternatives

T cells were isolated from human leukopaks using EasySep™ Human T Cell Isolation Kit (Catalog #17951), stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator (Catalog #10970), and cultured in either ImmunoCult™-XF medium supplemented with human recombinant IL-2 (rhIL-2) or other commercially available media. All commercial alternatives were prepared according to their respective manufacturer recommendations and supplemented with human AB serum or a serum derivative. The cells were analyzed at different time points for fold expansion relative to initial cell seeding density. ImmunoCult™-XF supported equal or higher expansion of total T cells compared to all commercial alternatives tested. T cells cultured in ImmunoCult™-XF underwent an average of 13-, 106-, and 876-fold expansion on days 5, 7, and 10, respectively. Data shown are mean ± SEM (n = 3 - 7 donors).

Figure 2. ImmunoCult™-XF Supports Equivalent or Greater T Cell Expansion than Other Commercial Alternatives

T cells were isolated from human leukopaks using EasySep™ Human T Cell Isolation Kit, stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator, and cultured in either ImmunoCult™-XF medium supplemented with rhIL-2 or commercially available media. All commercial alternatives were prepared according to their respective manufacturer recommendations and supplemented with human AB serum or a serum derivative. T cells were analyzed on day 10 for fold expansion relative to the initial cell seeding density and culture viability. Compared to all commercial alternatives tested, ImmunoCult™-XF showed similar or significantly higher expansion of total T cells (shown in bars) while supporting high viability (shown by data points) (> 95% viable on day 10). Data shown are mean + SEM (n = 3 - 7 donors). *p < 0.05, **p < 0.01 as determined by ANOVA using Dunnett's Multiple Comparison post-hoc test.

Figure 3. T Cells Expanded in ImmunoCult™-XF Maintain Similar Proportions of CD4+ and CD8+ Cells Following 10 Days of Culture Expansion

CD3+ T cells were isolated from human leukopaks using EasySep™ Human T Cell Isolation Kit, stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator, and cultured in ImmunoCult™-XF medium supplemented with rhIL-2. T cells were harvested on days (A) 0 and (B) 10 and analyzed by flow cytometry for the proportions of viable cells expressing CD4 and CD8. Data shown are from one representative donor; similar results were observed for the other 3 donors tested.

Figure 4. T Cells Expanded in ImmunoCult™-XF Express Intracellular IFN-Ɣ, TNF-α, and IL-4

T cells were isolated from human leukopaks using EasySep™ Human T Cell Isolation Kit, stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator, and cultured in ImmunoCult™-XF medium supplemented with rhIL-2. On day 10, T cells were harvested and analyzed for intracellular IFN-Ɣ, TNF-α, and IL-4 after stimulation with PMA and ionomycin for 4 hours, with Brefeldin A added after 2 hours. (A - C) IFN-Ɣ, TNF-α, and IL-4 production in CD4+ cells after stimulation; (D and E) IFN-Ɣ and TNF-α production in CD8+ cells after stimulation. Data shown are from one representative donor; similar results were observed for the other 3 donors tested. Unstimulated controls had < 0.5% IFN-Ɣ and TNF-α expression, or < 5% IL-4 expression (data not shown).