Achieve robust activation and expansion of mouse T cells in the absence of magnetic beads, feeder cells, or antigens.
The ImmunoCult™ Mouse T Cell Activator Kit’s gentle activation stimulus encourages high viability of activated T cells, which can be further expanded in common mouse T cell growth media. This kit consists of antibody complexes that bind to and cross-link CD3 and CD28 cell surface ligands, with the option to also target CD2, thereby providing the required primary and co-stimulatory signals for T cell culture and activation.
Optimal T cell activation can be achieved within 2 - 3 days using all 3 kit components (A, B, and C). Component A contains anti-mouse CD3 and CD28 antibodies for the necessary antigen and co-stimulatory activation signals. Component B contains anti-mouse CD2 antibodies for optional targeting. Component C helps to cross-link the antibodies in Component A and B. All reagents are highly stable, filter-sterilized and soluble, removing additional time-consuming plate coating steps during T cell culture, while allowing for a seamless integration into established workflows.
For efficient isolation of mouse T cells, the ImmunoCult™ Mouse T Cell Activator Kit can be combined with the EasySep™ workflow. It is compatible with kits such as EasySep™ Mouse T Cell Isolation Kit (Catalog #19851), EasySep™ Mouse CD4+ T Cell Isolation Kit (Catalog #19852), EasySep™ Mouse CD8+ T Cell Isolation Kit (Catalog #19853), EasySep™ Mouse Naive CD4+ T Cell Isolation Kit (Catalog #19765), EasySep™ Release Mouse CD4+CD304+ Regulatory T Cell Isolation Kit (Catalog #100-1570), and EasySep™ Mouse CD4 Positive Selection Kit II (Catalog #18952; Lung).
Cell Type T Cells, T Cells, CD4+, T Cells, CD8+, T Cells, Regulatory
Species Mouse
Application Activation, Cell Culture, Expansion
Brand ImmunoCult
Area of Interest Immunology, Cell Therapy Development
Data Figures
Figure 1. Activation of EasySep™-Isolated Mouse T Cells Stimulated with ImmunoCult™ Mouse T Cell Activator Kit
Mouse T cells were isolated using EasySep™ Mouse T Cell Isolation Kit, stimulated with ImmunoCult™ Mouse T Cell Activator Kit, and cultured in IMDM + FBS formulation. Following 3 days of culture, the mean ± SD frequency of CD25+ cells was 91.9 ± 5.1% (n = 11) or 99.9 ± 0.1% (n = 5), when stimulated with ImmunoCult™ Mouse CD3/CD28 T Cell Activator or ImmunoCult™ Mouse CD3/CD28/CD2 T Cell Activator, respectively. Stimulated mouse T cells maintained expression levels of CD25 throughout the 7-day culture period.
Figure 2. Robust Expansion of EasySep™-Isolated Mouse T Cells Can Be Achieved Following Stimulation with ImmunoCult™ Mouse T Cell Activator Kit
EasySep™-isolated mouse T cells were expanded with ImmunoCult™ Mouse T Cell Activator Kit in IMDM + FBS formulation over 7 days. The number of viable cells was assessed every 2 - 3 days, when fresh medium supplemented with IL-2 was added. No additional ImmunoCult™ Mouse T Cell Activator was added during the 7-day culture period. After 7 days in culture with ImmunoCult™ Mouse CD3/CD28 T Cell Activator or ImmunoCult™ Mouse CD3/CD28/CD2 T Cell Activator, stimulation resulted in a fold expansion of 23 ± 3.4 or 29.3 ± 4.8 (mean ± SEM, n = 6), respectively.
Figure 3. High Cell Proliferation Is Observed in EasySep™-Isolated T cells After Stimulation with ImmunoCult™ Mouse T Cell Activator
EasySep™-isolated mouse T cells were labeled with CFDA-SE (Catalog #75003), stimulated with ImmunoCult™ Mouse T Cell Activator Kit, and cultured in IMDM + FBS formulation. On Day 3, cells were harvested, stained with anti-mouse CD4 and CD8a antibodies, then measured by flow cytometry. Shown are CFDA-SE-labeled mouse T cells, gated on viable (A) CD4+ or (B) CD8a+ cells, cultured with no activator (top panel), with ImmunoCult™ Mouse CD3/CD28 T Cell Activator (middle panel), or with ImmunoCult™ Mouse CD3/CD28/CD2 T Cell Activator (bottom panel). Due to cell proliferation, the intensity of CFDA-SE signal is reduced by 50% for each cell division.
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.
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