ImmunoCult™ Mouse Th1 Differentiation Supplement

Supplement for the differentiation of mouse naïve CD4+ T cells into Th1 cells
概要
ImmunoCult™ Mouse Th1 Differentiation Supplement contains a combination of recombinant mouse cytokines and a blocking monoclonal antibody formulated to promote the differentiation of mouse naïve CD4+ T cells into Th1 cells. 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 Th1 function
• Development of procedures to expand and polarize naïve T cells toward a Th1 subset
Advantages
• Optimized for the induction of Th1 cells 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
Contains
• Recombinant mouse interleukin 2 (IL-2)
• Recombinant mouse interleukin 12 (IL-12)
• Rat anti-mouse interleukin 4 (anti-IL-4)
Subtype
Supplements
Cell Type
T Cells, T Cells, CD4+
Species
Mouse
Application
Cell Culture, Differentiation, Expansion
Brand
ImmunoCult
Area of Interest
Immunology
数据及文献

Data

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).

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