T Cell Isolation

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Flow Cytometry Application in CAR-T System

• Reference https//www.creative-biolabs.com/car-t/
  flow-cytometry-application-in-car-t-system.htm

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Background

• Flow cytometry is a widely used laser- or
  impedance-based method for analyzing the
  expression of cell surface and intracellular
  molecules, characterizing and defining different
  cell types in a heterogeneous cell population,
  assessing the purity of isolated subpopulations
  and analyzing cell size and volume. It allows
  simultaneous multi-parameter analysis of the
  physical and chemical characteristics of up to
  thousands of particles per second at single cell
  level.

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Flow Cytometry Application

• ? Patient's T Cell Isolation
• Flow cytometry analysis of isolated blood
  products ensures that a sufficient number of
  healthy and functional T cells being used for
  processing and re-infusion.
• ? CAR Expression Evaluation
• Assure that a sufficient number of viable T cells
  are expressing the specific CAR, and make sure
  that these cells show other phenotypic indicators
  of T cell functionality.
• ? CAR-T Cell Activation and Expansion
• Track T cell activation and expansion with cell
  proliferation and viability dyes in an easy way.

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Flow Cytometry Application

• ? CAR-T Cell Characterization
• analyze the phenotypic and subset composition of
  CAR-T cell populations.
• ? CAR-T Cell Function Monitoring
• monitor the persistence and functionality of
  these CAR-T cells as the evaluation of
  effectiveness of this treatment. It can be
  applied in functional cytokine production,
  cytokine release assay, immunohistochemistry
  analysis of CAR-T cell infiltration in tumor
  microenvironment, common immuno-activated or
  immunosuppressive markers expression etc.

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Flow Cytometry Application

• ? T Cell Exhaustion
• examine T cell exhaustion with markers including
  PD-L1 and FoxP3. Low exhausted phenotype ensures
  a more consistent and efficacious product.
• ? Tumor Regression Monitoring
• detect the frequency of target cell
  lysis/apoptotic cells, antitumor activity of
  CAR-T cell, clinical phenotypes of patients,
  disease diagnosis etc.

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• Referenc

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Thanks

• Reference https//www.creative-biolabs.com/car-t/
  flow-cytometry-application-in-car-t-system.htm