Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue

Summary: 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 micro...

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Main Authors: Maria Lamprou, Ana Krotenberg Garcia, Saskia Jacoba Elisabeth Suijkerbuijk
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724007408
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author Maria Lamprou
Ana Krotenberg Garcia
Saskia Jacoba Elisabeth Suijkerbuijk
author_facet Maria Lamprou
Ana Krotenberg Garcia
Saskia Jacoba Elisabeth Suijkerbuijk
author_sort Maria Lamprou
collection DOAJ
description Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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spelling doaj-art-02ba8ec7b51b4462a0cb3f59cbd9141a2025-01-21T04:13:19ZengElsevierSTAR Protocols2666-16672025-03-0161103575Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissueMaria Lamprou0Ana Krotenberg Garcia1Saskia Jacoba Elisabeth Suijkerbuijk2Division of Developmental Biology, Institute of Biodynamics and Biocomplexity, Department of Biology, Faculty of Science, Utrecht University, Padualaan 8, Utrecht 3584 CH, the Netherlands; Corresponding authorDivision of Developmental Biology, Institute of Biodynamics and Biocomplexity, Department of Biology, Faculty of Science, Utrecht University, Padualaan 8, Utrecht 3584 CH, the NetherlandsDivision of Developmental Biology, Institute of Biodynamics and Biocomplexity, Department of Biology, Faculty of Science, Utrecht University, Padualaan 8, Utrecht 3584 CH, the Netherlands; Corresponding authorSummary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724007408CancerMicroscopyOrganoids
spellingShingle Maria Lamprou
Ana Krotenberg Garcia
Saskia Jacoba Elisabeth Suijkerbuijk
Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
STAR Protocols
Cancer
Microscopy
Organoids
title Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
title_full Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
title_fullStr Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
title_full_unstemmed Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
title_short Protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
title_sort protocol for generating liver metastasis microtissues to decipher cellular interactions between metastatic intestinal cancer and liver tissue
topic Cancer
Microscopy
Organoids
url http://www.sciencedirect.com/science/article/pii/S2666166724007408
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