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CAR T Cell Characterization With Flow Cytometry
Adoptive immunotherapy is the transfer of immune cells into a patient for persistent anti-cancer treatment. Chimeric antigen receptors (CARs) cells comprise a type of adoptive therapeutic, with recombinant receptor constructs expressed in T cells to target cells expressing specific antigens. CAR T cells destroy tumor cells in certain B cell cancers and are making progress in eradicating solid tumors.
Research the next-generation of CAR therapeutics faster with flow cytometry. Learn about tools to detect T cell exhaustion and detect gene transcripts.
How it works: CAR T cells induce tumor cell death
CAR T cell flow cytometry workflow
Isolate T cells from donor sample
Stimulate T cells with antibody-coated Dynabeads magnetic beads for expansion
Express recombinant receptor constructs within T cells
Examine recombinant receptor expression with NGS or PrimeFlow assay
Assay for cell viability and functionality
Test for tumor regression in mouse models
Uses for flow cytometry during CAR T cell development
T cell activation and monitoring expansion
T cells do not proliferate without specific signals. Expand and test the quality of T cells for cell engineering with Dynabeads magnetic beads and cell health assays. Dynabeads magnetic beads isolate, activate, and expand T cells with clinical-grade beads coupled to primary and co-stimulatory signals anti-CD3 and anti-CD28 antibodies. This eliminates the need for feeder cells or manually adding cytokines for T cell stimulation. Then, track T cell expansion with cell proliferation and viability dyes.
Examining CAR T cell exhaustion
T cells including engineered CAR T cells can be inhibited by immunosuppressive molecules secreted in the tumor microenvironment. Complement immunohistochemistry analysis of CAR T cell–infiltrated tumor sections with flow cytometry of dissociated tumor cells. Count cells and examine multiple markers in one sample including common immunosuppressive markers IDO1, PD-L1, FoxP3, TDO, IL-10, and TGFβ.
Resources
Flow Cytometry Learning Center
Access flow cytometry educational resources for better experiment planning and execution.
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Flow Cytometry Support Center
Find technical support recommendations for your flow cytometry workflows, including tips for experimental setup and in-depth troubleshooting help.