Optimizing the viability, stability, and potency of Buffy coat isolated T cells for homologous dendritic cell co-cultures: A method for handling and preservation

Abstract

The utilization of T cells is becoming increasingly prominent in both clinical and research domains. However, the need to optimize preservation methodologies for extended periods of time remains unmet. To address this issue, we have developed a protocol for the handling and preservation of T cells that facilitates successful donor homologous co-cultures with dendritic cells (DCs), and preserves the cells for subsequent testing. Our method enhances experimental efficiency by reducing time and effort, and simplifying the use of T cells in mono or co-cultures. Our T cell handling and preservation methodology demonstrates the stability and viability of these cells in co-cultures, with viability exceeding 93% before and after liquid nitrogen preservation. Additionally, the preserved cells display no unspecific activation, as evidenced by the unaltered expression of the T cell activation marker CD25. The proliferation profile of preserved T cells used in DC-T cell co-cultures, stimulated by lipopolysaccharide (LPS)-activated DCs, attests to the potency and ability of these cells to interact and proliferate. These findings underscore the efficacy of our handling and preservation methodology in maintaining T cell viability and stability. Preserving donor T cells not only reduces the inconvenience of repeated blood donations but also enhances accessibility to a particular population of T cells for experimental or clinical applications, such as chimeric antigen receptor T cells.