ABSTRACT We describe a sensitive, robust, high-throughput method for quantifying the ability of metastatic tumor cells to colonize a secondary organ. Metastasis is the leading cause of death

in cancer patients, and successful colonization of the secondary organ is the rate-limiting step in the metastatic process; thus, experimental methods that can be used to interrogate the

key factors required for this critical step are of great importance. The experimental metastasis assay we detail here includes tail-vein injection of cancer cells into the mouse and

determination of the resulting secondary organ colonization, primarily in the lung, 10 d post dosing. This assay can be used to investigate factors that regulate metastatic colonization both

regulators of metastatic colonization. The experimental details discussed here include choosing of appropriate cell numbers, handling of the cells, selection of recipient animals and

injection techniques. Furthermore, we discuss key experimental design considerations, including the choice of the method used to determine metastatic burden and statistical analysis of the

results, as well as provide troubleshooting tips and identification of factors that contribute to experimental variability. Access through your institution Buy or subscribe This is a preview

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* Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS DISSECTING METASTASIS USING PRECLINICAL MODELS AND

METHODS Article 03 May 2023 INTEGRATING MODEL SYSTEMS AND GENOMIC INSIGHTS TO DECIPHER MECHANISMS OF CANCER METASTASIS Article 10 March 2025 EXPERIMENTAL AND SPONTANEOUS METASTASIS ASSAYS

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mouse screen data from studies to assess host regulation of metastatic colonisation. _Sci. Data_ http://dx.doi.org/10.1038/sdata.2017.129 (2017). Download references ACKNOWLEDGEMENTS This

work was supported by grants from Cancer Research UK (C20510/A13031 to D.J.A.), the Wellcome Trust (WT098051 to D.J.A.), ERC Combat Cancer (319661 to D.J.A.) and the National Institutes of

Health (U54HG004028 to N.A.K.). The authors thank the members of the Research Support Facility (Wellcome Trust Genome Campus) for their care of the mice, and M. Nohadani for histological

processing of the lung samples. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Experimental Cancer Genetics Group, Wellcome Trust Sanger Institute,, Hinxton, UK Anneliese O Speak, Agnieszka

Swiatkowska, Natasha A Karp, David J Adams & Louise van der Weyden * Quantitative Biology, Innovative Medicines and Early Development (IMED), AstraZeneca,, Cambridge, UK Natasha A Karp *

Cancer Research UK Edinburgh Centre, Institute of Genetics & Molecular Medicine (IGMM), Mark J Arends * University of Edinburgh, Edinburgh, UK Mark J Arends Authors * Anneliese O Speak

View author publications You can also search for this author inPubMed Google Scholar * Agnieszka Swiatkowska View author publications You can also search for this author inPubMed Google

Scholar * Natasha A Karp View author publications You can also search for this author inPubMed Google Scholar * Mark J Arends View author publications You can also search for this author

inPubMed Google Scholar * David J Adams View author publications You can also search for this author inPubMed Google Scholar * Louise van der Weyden View author publications You can also

search for this author inPubMed Google Scholar CONTRIBUTIONS L.v.d.W. and D.J.A. conceived the idea of screening mice from large-scale mouse phenotyping pipelines for metastatic

colonization. L.v.d.W., A.S. and A.O.S. performed the experiments. N.A.K. performed statistical analysis. M.J.A. performed histopathology analysis. L.v.d.W. and A.O.S. wrote the manuscript

with contributions from all authors. CORRESPONDING AUTHOR Correspondence to Louise van der Weyden. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial

interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY METHODS (PDF 431 KB) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Speak, A., Swiatkowska, A., Karp,

N. _et al._ A high-throughput _in vivo_ screening method in the mouse for identifying regulators of metastatic colonization. _Nat Protoc_ 12, 2465–2477 (2017).

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