The method is an approach to systematically study the impact of the nutrient microenvironment on the metabolism of effector CD8+ T cells, based on performing stable 13C isotope labeling measurements on in vitro-differentiated murine effector CD8+ T cells. Naive CD8+ T cells are isolated from mouse

Metabolomics and 13C tracer analysis are state-of-the-art techniques that allow determining the concentration of metabolites and the activity of metabolic pathways, respectively. Three dimensional (3D) cultures of cancer cells constitute an enriched in vitro environment that can be used to assay

We propose to generate three-dimensional tumoroids from the primary melanoma cell lines, as well as 3D organoids from the iPSc-derived neural crest stem cells. We will use the AggreWell system (STEMCELL Technologies) to generate uniform, size-controlled three-dimensional spheroids. After 5 days in