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 cells induce tumor cell death

CAR T cell flow cytometry workflow

Isolate T cells from donor sample

Isolate T cells from donor sample

Isolate T cells from donor sample

Stimulate T cells with antibody-coated Dynabeads magnetic beads for expansion

Express recombinant receptor constructs within T cells

Express recombinant receptor constructs within T cells

Examine recombinant receptor expression with NGS or PrimeFlow assay

Examine recombinant receptor expression with NGS or PrimeFlow assay

Assay for cell viability and functionality

Assay for cell viability and functionality

Test for tumor regression in mouse models

Test for tumor regression in mouse models


Uses for flow cytometry during CAR T cell development

T cell activation and monitoring expansion

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

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.