ImmunoCult Ms Treg Diff Suppl, 1mL

A cell culture supplement from STEMCELL Technologies, formulated to drive the differentiation of mouse naïve CD4+ T cells into regulatory T cells (Tregs). The supplement is a combination of recombinant mouse interleukin 2 (IL-2), recombinant human transforming growth factor beta (TGF-β), and all-trans retinoic acid (ATRA), supplied as a 100X concentrate that is added to culture medium at a 1 in 100 dilution.

For differentiation, flat-bottom tissue culture-treated cultureware is first coated with anti-mouse CD3 antibody, then naïve CD4+ T cells isolated from mouse splenocytes are cultured in RPMI 1640 Medium supplemented with fetal bovine serum, L-glutamine, HEPES buffer, non-essential amino acids, sodium pyruvate, and β-mercaptoethanol, together with anti-mouse CD28 antibody, for 6 days at 37°C and 5% CO₂. The protocol is optimized for naïve CD4+ T cells isolated from the spleen of C57BL/6 mice.

Under these Treg-polarizing conditions, the mean proportion of CD4+FOXP3+ cells is 91 ± 2% compared with 2 ± 0.4% in non-polarized cultures (p < 0.001; n = 14), and a 3.6-fold expansion of total nuclear cells is observed relative to non-polarized cultures (p < 0.001; n = 14). The supplement is stored at -20°C and is stable for 18 months from the date of manufacture on the label; once thawed it can be stored at 2 - 8°C for up to 1 month and should not be re-frozen.