Generation of vessel co-option lung metastases mouse models for single-cell isolation of metastases-derived cells and endothelial cells

Anne Cuypers; Laure-Anne Teuwen; Victoria L Bridgeman; Laura P M H de Rooij; Guy Eelen; Mieke Dewerchin; Anna Rita Cantelmo; Joanna Kalucka; Ann Bouché; Stefan Vinckier; An Carton; Ann Manderveld; Peter B Vermeulen; Andrew R Reynolds; Peter Carmeliet

doi:10.1016/j.xpro.2022.101691

Generation of vessel co-option lung metastases mouse models for single-cell isolation of metastases-derived cells and endothelial cells

STAR Protoc. 2022 Dec 16;3(4):101691. doi: 10.1016/j.xpro.2022.101691. Epub 2022 Sep 28.

Authors

Affiliations

¹ Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology (CCB), VIB, Department of Oncology, Leuven Cancer Institute, KU Leuven, 3000 Leuven, Belgium.
² Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology (CCB), VIB, Department of Oncology, Leuven Cancer Institute, KU Leuven, 3000 Leuven, Belgium; Translational Cancer Research Unit, GZA Hospitals Sint-Augustinus, 2610 Antwerp, Belgium; Center for Oncological Research, University of Antwerp, 2000 Antwerp, Belgium.
³ Tumour Host Interaction Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
⁴ Translational Cancer Research Unit, GZA Hospitals Sint-Augustinus, 2610 Antwerp, Belgium; Center for Oncological Research, University of Antwerp, 2000 Antwerp, Belgium.
⁵ Oncology R&D, AstraZeneca, Cambridge CB2 0AA, UK.
⁶ Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology (CCB), VIB, Department of Oncology, Leuven Cancer Institute, KU Leuven, 3000 Leuven, Belgium; Laboratory of Angiogenesis and Vascular Heterogeneity, Department of Biomedicine, Aarhus University, 8000 Aarhus, Denmark; Center for Biotechnology, Khalifa University of Science and Technology, Abu Dhabi 127788, United Arab Emirates. Electronic address: peter.carmeliet@kuleuven.be.

Abstract

Tumor vessel co-option, a process in which cancer cells "hijack" pre-existing blood vessels to grow and invade healthy tissue, is poorly understood but is a proposed resistance mechanism against anti-angiogenic therapy (AAT). Here, we describe protocols for establishing murine renal (RENCA) and breast (4T1) cancer lung vessel co-option metastases models. Moreover, we outline a reproducible protocol for single-cell isolation from murine lung metastases using magnetic-activated cell sorting as well as immunohistochemical stainings to distinguish vessel co-option from angiogenesis. For complete details on the use and execution of this protocol, please refer to Teuwen et al. (2021).

Keywords: Antibody; Cancer; Cell isolation; Model organisms; Single cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Disease Models, Animal
Endothelial Cells
Lung Neoplasms* / pathology
Mice
Neovascularization, Pathologic* / pathology