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A rapid and high-throughput T cell immunophenotyping assay for cellular therapy bioprocess using the Cellaca® PLX
Carolina Franco Nitta1, Mackenzie Pierce1, Jeanne Elia2
1Department of Consumables and Reagent Development, Revvity Health Sciences, Inc., Lawrence, MA 01843, USA.
Journal of Immunological Methods
|August 19, 2023
Summary
A new image cytometry system, Cellaca PLX, offers a faster and efficient alternative for cellular immunophenotyping and viability analysis. This method provides comparable results to traditional flow cytometry, streamlining workflows for cellular therapy development.
Area of Science:
- Cellular Therapy
- Immunology
- Biotechnology
Background:
- Chimeric antigen receptor (CAR) T and NK cells require phenotypic analysis during therapy development.
- Flow cytometry is a standard method for analyzing T cell populations (CD3, CD4, CD8).
- Image cytometry offers an alternative or orthogonal approach for cell-based assays.
Purpose of the Study:
- To demonstrate a rapid, high-throughput image cytometry method for cellular immunophenotyping and viability.
- To evaluate the Cellaca PLX system for analyzing critical quality attributes in cellular therapy samples.
- To compare the Cellaca PLX system's performance against traditional flow cytometry.
Main Methods:
- Utilized the Cellaca PLX image cytometry system for high-throughput analysis.
- Processed peripheral blood mononuclear cells (PBMCs) through RBC lysis and CD3 enrichment.
- Stained samples with specific surface marker kits (Hoechst/CD3/CD4/CD8 or Hoechst/CD3/CD8/RubyDead Dye).
- Performed side-by-side comparisons with three different flow cytometers for validation.
Main Results:
- The Cellaca PLX system demonstrated faster and more efficient acquisition and analysis of cell viability and populations.
- Results from the Cellaca PLX were highly comparable to those obtained from flow cytometry.
- The image cytometry method effectively assessed cell identity, viability, and other biological functions.
Conclusions:
- The Cellaca PLX Image Cytometer serves as a rapid alternative or orthogonal method for PBMC immunophenotyping.
- This system can streamline workflows, enabling quicker downstream processing of valuable patient samples.
- The technology supports critical quality attribute assessment for cellular therapeutic products, aligning with ISO standards.
