Figure 1. ImmunoCult™ Mouse Th1 Differentiation Supplement Produces CD4+IFNγ+T-Bet+ Cells Under Th1 Polarizing Conditions

Naïve CD4+ T cells were isolated from mouse splenocytes using the EasySep™ Mouse Naive CD4+ T Cell Isolation Kit (Catalog #19765), activated with plate-bound anti-CD3 and soluble anti-CD28 and cultured in medium alone (Non-polarized cultures), or medium supplemented with ImmunoCult™ Mouse Th1 Differentiation Supplement (Polarized cultures ) for 5 days. Cells were subsequently stimulated with PMA/Ionomycin, in conjunction with brefeldin A treatment, followed by staining with anti-CD4, anti-IFNγ, anti-T-bet and a viability dye and analyzed by flow cytometry. Shown is the expression of IFNγ and T-bet, back-gated on viable, CD4+ cells from non-polarized (A) or polarized cultures (B). The mean proportion of CD4+IFNγ+T-Bet+ cells is significantly higher in cells cultured in ImmunoCult™ Mouse Th1 Differentiation Supplement (44 ± 6%) compared to non-polarized cells (2 ± 1%) (p < 0.001; n = 13). Data from experimental groups were compared using a paired T-test.

Figure 2. ImmunoCult™ Mouse Th1 Differentiation Supplement Expands Total Nucleated Cells Under Th1 Polarizing Conditions

Naïve CD4+ T cells were isolated from mouse splenocytes using the EasySep™ Mouse Naive CD4+ T Cell Isolation Kit (Catalog #19765), activated with plate-bound anti-CD3 and soluble anti-CD28 and cultured in medium alone, or medium containing ImmunoCult™ Mouse Th1 Differentiation Supplement for 5 days. A 4.5-fold expansion of total nuclear cells (TNC) ensues in cultures polarized with ImmunoCult Mouse Th1 Differentiation Supplement (purple column), which is significantly higher compared to non-polarized cultures (grey column). (Data represent the mean ± SEM, n = 11 - 20, *p < 0.05. Data from experimental groups were compared using a paired T-test).