ImmunoCult™ Mouse Treg Differentiation Supplement

Supplement for the differentiation of mouse naïve CD4+ T cells into regulatory T cells (Tregs)

ImmunoCult™ Mouse Treg Differentiation Supplement

Supplement for the differentiation of mouse naïve CD4+ T cells into regulatory T cells (Tregs)

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Supplement for the differentiation of mouse naïve CD4+ T cells into regulatory T cells (Tregs)
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Product Advantages


  • Optimized for the induction of Tregs from naïve CD4+ T cells isolated from the spleen of a C57BL/6 mouse

  • Supplied as a 100X concentrate; after thawing and mixing, the tube contents can be added directly to medium

Overview

ImmunoCult™ Mouse Treg Differentiation Supplement contains a combination of recombinant mouse and human cytokines and a small molecule optimized to promote the differentiation of mouse naïve CD4+ T cells into regulatory T cells (Tregs). This supplement is intended for use with RPMI 1640 Medium (Catalog #36750) containing fetal bovine serum and other additives, as well as anti-mouse CD3 and anti-mouse CD28 monoclonal antibodies as activating agents.

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
Contains
• Recombinant mouse interleukin 2 (IL-2)
• Recombinant human transforming growth factor beta (TGF-β)
• All-trans retinoic acid (ATRA)
Subtype
Supplements
Cell Type
T Cells, T Cells, CD4+, T Cells, Regulatory
Species
Mouse
Application
Cell Culture, Differentiation, Expansion
Brand
ImmunoCult
Area of Interest
Immunology

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.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
10957
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
10957
Lot #
All
Language
English

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.