Culture surface influence on T-cell phenotype and function
Issue title: Selected articles of the 32nd Annual Conference of the German Society for Clinical Hemorheology and Microcirculation, Dresden, Germany, 24 – 25 May, 2013
Affiliations: Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany | Institute of Biomaterial Science, Helmholtz - Zentrum Geesthacht, Teltow, Germany | Medical Department, Division of Nephrology and Internal Intensive Medicine, Charité - Universitätsmedizin Berlin, Berlin, Germany | Institute of Medical Immunology, Charité - Universitätsmedizin, Berlin, Germany
Note: [] Corresponding author: Petra Reinke, Medical Department, Division of Nephrology and Internal Intensive Medicine, Charité - Universitätsmedizin Berlin, Charité Campus Virchow-Klinikum, Augustenburger Platz 1, 13353 Berlin, Germany. E-mail: [email protected]
Abstract: When dealing with T lymphocyte culture there is currently very less information available about the interaction between T-cells and the culture system. In this study we look at the influence of the culture chamber on T-cell proliferation in two main aspects of the culture system, namely: culture chamber material and geometry. The study was carried out using unique polymeric closed cell culture inserts, which were processed via injection moulding from polystyrene (PS), polycarbonate (PC), polyetherurethane (PEU), polystyrene-co-acrylonitrile (PSAN) and polyetherimide (PEI). Furthermore culture chamber geometry was studied using commercially available 24, 12 and 6-well plates prepared from tissue culture plastic (TCP). For T lymphocyte stimulation two methods were used involving either EBV peptide pools or MACS iBead particles depending on the experiment performed. Culture was done with 1645 RPMI medium supplemented with foetal calf serum, penicillin, streptomycin and rhIL-2. We found four materials out of five we tested (PS, PC, PSAN and PEI) exhibited similar fold expansions with minimal influence on proportions of CD4 and CD8, while PEU had a negative influence on T cell growth along with adversely affected CD4/CD8 proportions. Changes in the geometry of TCP had no effect on T cell growth or maturation rather the size of geometry seems to have more influence on proliferation. T-cells appear to prefer smaller geometries during initial stages of culture while towards the end of the culture size becomes less significant to cell proliferation. The parameters tested in this study have significant influences on T-cell growth and are necessary to consider when designing and constructing expansion systems for antigen specific T lymphocytes. This is important when culturing T-cells for immunotherapeutic applications where antigen specificity, T-cell maturation and function should remain unaffected during culture.
Keywords: T-cell culture, immunotherapy, material testing, geometry
