Protocol for biomimetic tumoroid models by plastic compression using centrifugation

Here, we present a protocol for engineering biomimetic tumoroid models by plastic compression using centrifugation. We describe steps for generating multi-compartment tumor-stroma models by mixing cells into a collagen hydrogel crosslinked at 37°C and centrifuging the hydrogel. We then detail procedures for generating compartmentalized models and encapsulating the final layered hydrogel containing a 96-well tumor mass in a 24-well stroma. This protocol increases collagen density and improves mechanical properties of collagen hydrogels.