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However, these formulations may not recapitulate all the nutritional environmental cues required to induce and sustain robust expansion specific to T cells necessitating their supplementation with human AB serum. This has led to the generation of culture media that recapitulate many physiological characteristics of human plasma such as Human Plasma-like Medium (HPLM) 21 and others 22. However, recent evidence has indicated that cells maintained under these non-physiological conditions activate different signaling and metabolic pathways compared to analogous cells generated in vivo 20. Improvements in formulations that incorporate defined serum components have led to media formulations such as X-VIVO 15 and AIM-V. The first synthetic complex medium used for in vitro expansion of lymphocytes was RPMI 1640 18, and is widely used in the Rapid Expansion Protocol (REP) of TILs 19. At the present time, there has yet to be a universally accepted formulation to manufacture T cells making it a major challenge not only to cross-compare different clinical trials, but also to understand the metabolic parameters that may be responsible for the behavior of T cells post-transfer. A recent study identified medium-dependent transcriptional responses in several metabolic pathways during early activation, which may be crucial in programming T cells to a committed phenotype 13– 17. Culture methods that maintain metabolic profiles favoring central memory phenotypic subsets have been shown to highly correlate with improved clinical outcomes 12. Another critical factor is the complex cell manufacturing process that can generate products with undesirable metabolic phenotypes and ultimately hamper in vivo functionality 9– 11. These include but are not limited to tumor antigen escape, restricted intraepithelial trafficking, limited persistence and an inhospitable tumor microenvironment (TME) 6– 8. However several barriers exist that impede the efficacy of cell based therapies targeting solid tumors 3– 5. In particular, CAR T cell therapy has achieved exceptional response rates in hematological cancers 1, 2. These therapies involve isolating and expanding autologous human tumor-infiltrating lymphocytes (TIL) or genetically modified chimeric antigen receptor (CAR) T cells. Thus, ex vivo T cell expansion conditions have profound impacts on metabolism and function.Ĭell-based immunotherapies are garnering considerable attention for their promise in treating human cancers. In one case, culturing in ascites resulted in increased glucose uptake which was insufficient to rescue T cell function. Furthermore, T cell products cultured in ascites from ovarian cancer patients displayed suppressed mitochondrial activity and function irrespective of the ex vivo expansion media. However, adoption of these metabolic phenotypes was uncoupled to T cell function. T cells adapted their metabolism to match their media expansion condition as shown by glucose and glutamine uptake, and patterns of glucose isotope labeling. The comparison of five different culture media used for clinical T cell expansion revealed unique optima based on different output variables including proliferation, differentiation, function, activation and mitochondrial phenotypes.
INCREASE FONT SIZE TEXMACS MAC OS
See our step-by-step instructions for increasing the text size on Mac OS overall, changing the icon font size and enlarging the finder sidebar's font.Ex vivo expansion conditions used to generate T cells for immunotherapy are thought to adopt metabolic phenotypes that impede therapeutic efficacy in vivo. The simplest involves navigating to System Preferences->Displays, selecting the Scaled resolution option and picking a lower resolution than the default. There are several ways to increase the font size in Mac OS X.