ImmunoCult™ Mouse Treg Differentiation Supplement
Supplement for the differentiation of mouse naïve CD4+ T cells into regulatory T cells (Tregs)
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Overview
This product is recommended for:
• Research into the regulation of mouse naïve T cell differentiation and Treg function
• Development of procedures to expand and polarize naïve T cells toward a Treg subset
• Recombinant human transforming growth factor beta (TGF-β)
• All-trans retinoic acid (ATRA)
Data Figures

Figure 1. ImmunoCult™ Mouse Treg Differentiation Supplement Produces CD4+CD25+FOXP3+ Cells Under Treg 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-polarizing cultures), or in medium supplemented with ImmunoCult™ Mouse Treg Differentiation Supplement (Polarizing cultures) for 6 days. Cells were subsequently stained with anti-CD4, anti-CD25, anti-FOXP3 and a viability dye and analyzed by flow cytometry. Shown is the expression of CD25 and FOXP3 back-gated on viable, CD4+ cells from non-polarized (A) or polarized cultures (B). The mean proportion of CD4+FOXP3+ cells is significantly higher in cells cultured in ImmunoCult™ Mouse Treg Differentiation Supplement (91 ± 2%) compared to non-polarized cells (2 ± 0.4%) (p < 0.001; n = 14). Data from experimental groups were compared using a paired T-test.

Figure 2. ImmunoCult™ Mouse Treg Differentiation Supplement Expands Viable CD4+ Cells Under Treg 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 Treg Differentiation Supplement for 6 days. A 3.6-fold expansion of total nuclear cells (TNC) ensues in cultures polarized with ImmunoCult Mouse Treg Differentiation Supplement (purple column), which is significantly higher compared to non-polarized cultures (grey column). (Data represent the mean ± SEM, n = 14, ***p < 0.001. Data from experimental groups were compared using a paired T-test).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (4)
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.