Description

Human CD3+ T cell Potential Application Summary

• Flow cytometry-based binding studies for target binding and cross-reactivity
• Large-scale potency assays to compare a panel of test articles for the ability to elicit T cell responses
• Adoptive transfer into immunodeficient recipient mice to support the in vivo evaluation of activity with therapeutic test articles

Expansion of Human CD3+ T cells

These human CD3+ T cells are meticulously expanded over a multi-day period using a combination of cytokines, expansion medium, and high quality T cell activation beads. The expansion process takes place in a controlled environment to ensure optimal conditions for cell growth and retained function.

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Figure 1: T cell phenotype is maintained post-expansion. (a) CD3+ purity is maintained after expansion. (b) CD4+ T cell frequency decreases after expansion with a concordant increase in CD8+ T cell frequency. Data shows one representative sample.

Validation of Expanded Human CD3+ T cells

Our expanded human CD3+ T cells have undergone rigorous testing to validate their exceptional quality and functional ability as reflected by their capacity for robust proliferation, target cell killing, and cytokine production, in vitro. These cells may be used for various applications, including mechanistic research, therapeutic development, and innovative cellular assays.

Proliferation

Figure 2: CD8+ T cells within expanded human CD3+ T cells proliferate in response to stimulation. Expanded human T cells were stimulated with activation beads for 96 hours. Flow cytometry was used to assess the frequency of Ki-67+ (a marker of proliferation) cells within CD8+ T cells. Data shows the average (mean) of technical triplicates with SEM error bars.

Cytokine Release

Figure 3: Expanded human CD3+ T cells secrete cytokines upon activation. Expanded human T cells were stimulated with activation beads for 24 hours. Cell-free supernatant was harvested, and cytokine secretion was quantified via cytometric bead array. In response to stimulation, expanded T cells secrete IL-2, IL-10, TNF-α, and IFN-γ. Data shows the average (mean) of technical triplicates with SEM error bars.

T cell Dependent Cellular Cytotoxicity (TDCC)

Figure 4: Expanded human CD3+ T cells exhibit cytotoxicity toward tumor target cells. Expanded human CD3+ T cells were co-cultured with fluorescently-labeled Ramos cells and treated with either a human-reactive bispecific T cell engager antibody (CD3xCD19) or isotype control. Ramos target cell cytotoxicity was measured on a flow cytometer via viability dye incorporation. Data shows the average (mean) of technical triplicates, normalized with background subtraction, with SEM error bars.

Cryopreservation and storage

Expanded human CD3+ T cells are carefully cryopreserved using iQ Biosciences’ cryopreservation protocol that ensures high viability (> 90%) upon thaw.

Cells should be stored at < -120 °C once they are received, such as within a liquid nitrogen tank (vapor phase).