Protocol for establishing a coculture with fibroblasts and colorectal cancer organoids
Svenja K. Wallisch, Sylvia Karin Neef, Lukas Denzinger, Dina Mönch, J. Koch, Julia Marzi, Thomas E. Mürdter, Nicole Janssen
Abstract
The tumor microenvironment is essential for mediating drug resistance and tumor progression. Here, we present a coculture system, which enables drug testing of colorectal cancer organoids and fibroblasts without additional matrix components such as Matrigel or basement membrane extracts. First, we describe steps to use a readout for high-throughput drug testing using a luminescence-based viability assay. Second, we detail a readout that uses flow cytometry to distinguish toxic effects on either colorectal cancer organoids or fibroblasts.
Topics & Concepts
OrganoidMatrigelColorectal cancerCancer-Associated FibroblastsFlow cytometryTumor microenvironmentCancer researchCancer cellBiologyChemistryCell biologyCancerTumor cellsMolecular biologyAngiogenesisGeneticsCancer Cells and Metastasis3D Printing in Biomedical ResearchChemical Reactions and Isotopes