» Articles » PMID: 39836518

Protocol for Generating Liver Metastasis Microtissues to Decipher Cellular Interactions Between Metastatic Intestinal Cancer and Liver Tissue

Overview
Journal STAR Protoc
Publisher Cell Press
Date 2025 Jan 21
PMID 39836518
Authors
Affiliations
Soon will be listed here.
Abstract

Cell competition is a quality control mechanism that promotes elimination of suboptimal cells relative to fitter neighbors. Cancer cells exploit these mechanisms for expansion, but the underlying molecular pathways remain elusive. Here, we present a protocol for generating matrix-free microtissues recapitulating cellular interactions between intestinal cancer and hepatocyte-like cells using microscopy or transcriptomics/proteomics. We describe steps for generating and differentiating liver progenitor organoids and microtissue formation. We then detail procedures for immunofluorescence staining, mounting microtissues, and sorting cells. For complete details on the use and execution of this protocol, please refer to Krotenberg Garcia et al..

References
1.
Huch M, Dorrell C, Boj S, van Es J, Li V, van de Wetering M . In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature. 2013; 494(7436):247-50. PMC: 3634804. DOI: 10.1038/nature11826. View

2.
Kaufmann A, Mickoleit M, Weber M, Huisken J . Multilayer mounting enables long-term imaging of zebrafish development in a light sheet microscope. Development. 2012; 139(17):3242-7. DOI: 10.1242/dev.082586. View

3.
Fumagalli A, Suijkerbuijk S, Begthel H, Beerling E, Oost K, Snippert H . A surgical orthotopic organoid transplantation approach in mice to visualize and study colorectal cancer progression. Nat Protoc. 2018; 13(2):235-247. DOI: 10.1038/nprot.2017.137. View

4.
Kanton S, Pasca S . Human assembloids. Development. 2022; 149(20). DOI: 10.1242/dev.201120. View

5.
Broutier L, Andersson-Rolf A, Hindley C, Boj S, Clevers H, Koo B . Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat Protoc. 2016; 11(9):1724-43. DOI: 10.1038/nprot.2016.097. View