3D Ewing Sarcoma Modeling for Metabolic Studies.

Three-dimensional (3D) in vitro models provide a physiologically relevant platform that more accurately recapitulates tissue architecture, cell-cell and cell-extracellular matrix interactions, and the physical microenvironment than traditional two-dimensional (2D) cultures. In particular, these models reproduce metabolic gradients, including oxygen and nutrient availability, enabling more precise investigations of cellular metabolism. This chapter describes two methods to grow Ewing sarcoma cell lines as 3D spheroids using physiological media, as well as the collection of spheroids in bulk or individually for different downstream analyses, particularly those related to metabolic studies.