Physiologix™ Serum Replacement and NB-ROC™ T Cell Culture Media Combination Shows Enhanced T-Cell Transduction Efficiency for CAR-T Therapy

CD19 Chimeric Antigen Receptor (CAR-T) T cell therapy has shown a great potential for the treatment of B-cell malignancies and lymphoma and an increased number of studies are in progress for its translation to solid tumors. Success of this treatment is unfortunately limited by multiple factors such as donor-to-donor variability, patients' response to the therapy, and manufacturing-related issues. After leukapheresis, isolated T cells are manipulated under artificial conditions to be transfused back to the patient. Since these steps are conducted outside of the cell's natural surroundings, maintaining cell viability and their ability to proliferate without exhaustion are pillars for the success of the therapy. Thus, using the appropriate reagents is crucial for the treatment to be efficient, which will subsequently affect the quality of the end product and potentially increase the number of treated patients. Cell culture media is a critical factor to consider when improving the process as it is intimately linked to cell function and metabolism. In this study, we tested a new cell culture media, NB-ROC™, developed specifically to support activated T cell proliferation and lentivirus transduction. Based on our previous data generated with the University of Pennsylvania on T cells, we supplemented NB-ROC™ with 2% Physiologix™ XF (Human Growth Factor Concentrate (hGFC), a cGMP, xeno-free media supplement made for stem cells and T cells). Compared to CTS™ OpTmizer™ T-Cell Expansion, NB-ROC™ shows on Dynabeads-activated T cells a maintained proliferation rate and population doubling with a preserved phenotype, determined through the evaluation of surface expression markers such as PD-1, CCR-7, and CD45-RO over a period of 7 days. Lentivirus transduction data is more compelling as it shows an increased level by 20% compared to OpTmizer™ supplemented with Human Serum. Our data also confirms previous observations on Physiologix™, as supplementing CTS™ OpTmizer™ with 2% Physiologix™ shows significant transduction efficiency improvement compared to when it is supplemented with 5% Human Serum. All in all, our data shows that NB-ROC™, as serum-free media, combined with Physiologix™ offers a very promising alternative for an improved T cell transduction efficiency with a maintained phenotype and proliferation rate on both 6-well plate and G-Rex model. This set of data holds the promise for enhancing transduction efficiency for CAR-T therapy which will help ameliorate the process and bring more success to the field.

Ayari:Nucleus Biologics: Current Employment, Current holder of stock options in a privately-held company. Maradze:Nucleus Biologics: Current Employment, Current holder of stock options in a privately-held company. Kadota:Nucleus Biologics: Current Employment, Current holder of stock options in a privately-held company. O'Connor:Nucleus Biologics: Consultancy, Membership on an entity's Board of Directors or advisory committees.