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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).
Contains
Anti-mouse CD3 monospecific antibody
Anti-mouse CD28 monospecific antibody
Anti-mouse CD2 monospecific antibody
Antibody crosslinking complex
Subtype
Complete Kits
Cell Type
T Cells, T Cells, CD4+, T Cells, CD8+, T Cells, Regulatory
Figure 1. ImmunoCult™ Mouse T Cell Activator Kit Supports High Viability of Activated T Cells
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, and 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 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 CD4+ (A) or CD8a+ (B) 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.
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