# Protocol for biomimetic tumoroid models by plastic compression using centrifugation

## TL;DR

This paper introduces a method to create realistic tumor models using centrifugation to compress collagen gels, improving their mechanical properties for research.

## Contribution

A novel protocol for creating multi-compartment tumoroid models using plastic compression via centrifugation.

## Key findings

- Centrifugation increases collagen density in hydrogels.
- The method allows embedding tumor masses into stromal compartments.
- The protocol is scalable for high-throughput applications.

## Abstract

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.

•Instructions for seeding cells, cultured in 2D, within a collagen I hydrogel•Steps for centrifuging collagen hydrogels to remove fluid, producing dense collagen gels•Guidance for embedding a tumor mass into a relevant stromal compartment•Steps for centrifuging a complex collagen gel, making it ready for downstream applications

Instructions for seeding cells, cultured in 2D, within a collagen I hydrogel

Steps for centrifuging collagen hydrogels to remove fluid, producing dense collagen gels

Guidance for embedding a tumor mass into a relevant stromal compartment

Steps for centrifuging a complex collagen gel, making it ready for downstream applications

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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.