Characterizing the health of transduced CAR-T cells using high-throughput image cytometry

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B938
Abstract ID: 6109

Presenting Author:
Yongyang Huang , Senior Scientist at Revvity Hlth. Sci.

Abstract:

Chimeric antigen receptor (CAR)-T cell therapy is one of the novel cellular therapeutic options for cancer patients. In this work, we developed a rapid cell viability and apoptosis detection method to characterize the cell health and recovery of SubT1 cells after the plasmid-based electroporation process to create CAR expression cells using the Cellaca® PLX Image Cytometer. Viability measurement of transduced SupT1 cells was performed by staining with acridine orange/propidium iodide (AO/PI). In addition, early and late-stage apoptosis were detected by staining transduced SupT1 cells with Annexin V/PI and Caspase3/RubyDead, respectively. Results showed that the introduction of plasmids, other than the electroporation process itself, may induce 20 – 30% additional cell death in the process, indicated by viability results measured on Day 1 post transduction. Among all the Annexin V or Caspase3 positive transduced SupT1 cells on Day 1 (22 – 34%), 70 - 80% of these cells were double positive in both Annexin V and PI, or both Caspase3 and RubyDead, and only 20 – 30% of them were in the early-stage/late-stage apoptosis. Subsequently, the SupT1 cells were quickly recovered with their viabilities increased to >90% on Day 3. With such capability of rapid detection of cell viability and apoptosis, Cellaca® PLX Image Cytometer can potentially support the characterization of factors that affects the quality of CAR-T cells in the manufacturing process.