Figure 1.Typical EasySep™ Mouse T Cell Isolation Profile

Starting with mouse splenocytes, the T cell content (CD3+CD19-) of the isolated fraction is 96.6 ± 2.0% (mean ± SD), using the purple EasySep™ magnet.

Figure 2.Cell Isolation Protocol Lengths

Typical time taken (in minutes) to isolate cells using select EasySep™ kits.

Figure 3.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 (Catalog #19851), stimulated with ImmunoCult™ Mouse T Cell Activator Kit (Catalog #100-1572), 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 4.Robust Expansion of EasySep™-Isolated Mouse T Cells Can Be Achieved Following Stimulation with ImmunoCult™ Mouse T Cell Activator Kit

Mouse T cells isolated using EasySep™ Mouse T Cell Isolation Kit (Catalog #19851) were expanded with ImmunoCult™ Mouse T Cell Activator Kit (Catalog #100-1572) 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 5.High Cell Proliferation is Observed in EasySep™-Isolated T cells After Stimulation with ImmunoCult™ Mouse T Cell Activator

Mouse T cells isolated using EasySep™ Mouse T Cell Isolation Kit (Catalog #19851) were labeled with CFDA-SE (Catalog #75003), stimulated with ImmunoCult™ Mouse T Cell Activator Kit (Catalog #100-1572), 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.