Automated PBMC Isolation and T Cell Wash and Concentration by the CTS Rotea System

Successful processing and manufacturing in cell and gene therapy workflows are essential to the efficacy of the product. Autologous cell and gene therapy workflows involve isolating cells from an individual, engineering the cells, expanding and concentrating them, and infusing them back into the patient. Certain steps in these workflows could benefit from optimized automation to decrease hands-on time and the cost of the cell manufacturing process.

In this application note, we present the Gibco™ Cell Therapy Systems™ (CTS™) Rotea™ Counterflow Centrifugation System, a closed cell processing system developed for small-batch cell therapy manufacturing. We demonstrate the efficacy of the CTS Rotea system for isolating peripheral blood mononuclear cells (PBMCs) from leukapheresis products, and its suitability for washing and concentrating T cells—two common processes that are central in many cell therapy workflows.

We present data supporting the ability of the CTS Rotea system to isolate, wash, and concentrate various cell types with no phenotypic change and no loss in recovery or viability. We show that PBMCs isolated from a leukapheresis product had recovery and viability comparable to PBMCs manually isolated using a density gradient medium, with no change in phenotype or cell type composition. T cells isolated from PBMCs were successfully activated with Dynabeads magnetic beads, expanded, and characterized after processing with the CTS Rotea system. Expanded primary T cells washed and concentrated using the CTS Rotea system showed high recovery, cell concentration, and viability with no change in phenotype across various output volumes. The flexibility and efficiency of the system and user-programmable software allows it to be incorporated into multiple steps of various cell therapy workflows.