Figure 1. ImmunoCult™ Mouse Th2 Differentiation Supplement Produces CD4+GATA3+IL-4+ Cells Under Th2 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 in medium supplemented with ImmunoCult™ Mouse Th2 Differentiation Supplement (Polarizing cultures) for 6 days. Cells were subsequently stimulated with PMA/Ionomycin, in conjunction with monensin treatment, followed by staining with anti-CD4, anti-IL-4, anti-GATA3 and a viability dye and analyzed by flow cytometry. Shown is the expression of GATA3 and IL-4 back-gated on viable, CD4+ cells from non-polarized (A) or polarized cultures (B). The mean proportion of CD4+IL-4+GATA3+ cells is significantly higher in cells cultured in ImmunoCult™ Mouse Th2 Differentiation Supplement (25 ± 3%) compared to non-polarized cells (4 ± 1%) (p < 0.001; n = 10). Data from experimental groups were compared using a paired T-test.

Figure 2. ImmunoCult™ Mouse Th2 Differentiation Supplement Expands Total Nucleated Cells Under Th2 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 Th2 Differentiation Supplement for 6 days. A 62-fold expansion of total nuclear cells (TNC) ensues in cultures polarized with ImmunoCult Mouse Th2 Differentiation Supplement (purple column), which is significantly higher compared to non-polarized cultures (grey column). (Data represent the mean ± SEM, n = 11, ***p < 0.001. Data from experimental groups were compared using a paired T-test).