Evidence of Improved Transduction and Preservation of Relevant T Cell Subpopulations Using Different Media Conditions: Towards a Novel CAR-T Media Formulation

9 May 2019
A Poster About Evidence of Improved Transduction & Preservation of Relevant T-Cell Subpopulations

The field of cell therapy is shifting away from animal origin components used in the culture or preservation of cells and replacing them with human or chemically defined components. However, this shift often leads to diminished cell proliferation and functionality due to the lack of physiologically relevant proteins. Therefore, there is an important need for generation of media supplements to optimize cell therapy. Physiologix XF Human Growth Factor Concentrate is a cGMP, xeno-free media supplement that can be used as a replacement to supplements such as human serum.

In this study, T cells were isolated from blood and cultured for 14 days in OpTmizer or X-VIVO 15 supplemented with 5% human serum or 2% Physiologix. Population doublings and cell size were defined, as well as CD4/CD8 and memory markers (CCR7 and CD45RO) to assess phenotype. We detected an increase in naïve and central memory populations in 2% Physiologix compared to human serum across all tested basal media. These populations have been described as highly relevant for the outcome of patients undergoing CAR-T cell therapy. We compared the transduction efficiency of a lentiviral-GFP in OpTmizer, X-VIVO15 or RPMI, supplemented with 2% Physiologix or 5% human serum at different multiplicities of infection (MOIs). Higher transduction efficiency was observed when using 2% Physiologix at a high MOI of 4 and was conserved across all dilutions including a MOI of <1.

These results indicate that Physiologix can be used to optimize the generation of relevant T cell subpopulations and maximize transduction efficiency even at a low MOI, thus increasing the efficiency of vector usage. These results may lead to better clinical efficacy while reducing overall costs.

Download Guide

Share this article

Asma Ayari, Ph.D.
Dr. Asma Ayari is the Director of Research and Development at Nucleus Biologics. Dr. Ayari has a deep understanding of how complex cellular systems are affected by small changes in the chemical composition of the microenvironment and has extensive experience in media analysis, testing, and formulation for cell and gene therapy. She is a subject matter expert on cell culture media solutions, primarily focusing on media formulation design and development. Dr. Ayari holds a M.S. in Microbiology from University of Tunis and a Ph.D. in Virology, Immunology, and Metabolism from Universite de Lille in France.