The ex vivo expansion of T cells is a critical process in bio-manufacturing of adoptive cell therapies. Recent clinical studies show a correlation between persistence of subsets of functional memory T cells, including T memory stem cells (TSCM), central memory T cells (TCM) and other less differentiated T cell subsets, and long-term anti-tumor responses in patient outcomes. Technologies that can robustly and reliably provide high-content immunophenotypic and functional data are therefore critical for the rapid and robust development of new therapies.
In an on-demand SelectScience webinar, Dr. John O'Rourke explores how a rapid multiparametric assay was developed and evaluated using the Intellicyt iQue3, an advanced flow cytometry platform, to address the need to profile T cell memory subsets and functional cytokine release.
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Q: Can this assay be measured in a 384-well plate format?
A: Yes, we have performed this assay using 5 µl of cell supernatant mixture in a 384-well plate without any deviation from the workflow.
Q: Can the ForeCyt Software be adapted to one-sample platforms, such as FacScalibur?
A: No, it's not a third-party software, it can only be used with the iQue3 instrument and is part of our integrated platform system.
Q: Have you used the cell and bead workflow for other applications?
A: Yes, we have used a similar workflow to assess human T cell activation where we measured early and late markers of activation, as well as cell proliferation and cytokine release.
Q: Have you measured other cytokines in this assay?
A: Yes, in this application we have measured and validated up to eight cytokines simultaneously using this workflow including (IL-2, 6, 10, 13, 17A, GM-CSF, IFNg, & TNF).
Q: Why was CD28 chosen as an activation marker?
A: We used anti-CD28, in conjunction with anti-CD3, on beads to activate T cells, as this is a standard protocol for T cell activation.
Q: How can I get a demo for some hands-on experience?
A: For more information on a demo, please contact us at firstname.lastname@example.org
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