AF Chambers

Agnieszka Swiatkowska

Anneliese O Speak

B Lucke

C Khanna

CP Klerk

David J Adams

DR Hurst

G Edel

IJ Fidler

IR Hart

JB Seo

JP Quigley

JT Wessels

KJ Luzzi

KW Brunson

L van der Weyden

Louise van der Weyden

M Bouvet

M Nogawa

MA Chekmareva

Mark J Arends

MP Rodero

NA Karp

Natasha A Karp

P Orosz

PJ Curran

PS Steeg

RM Hoffman

S Heilmann

S Koop

S Paget

SF Goldberg

TH Kuo

YG Park

ZM Zhao

English

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 at the tumor-cell-intrinsic level (via manipulation of the tumor cells before injection) and at the tumor-cell-extrinsic level (such as the tissue microenvironment, via the use of genetically modified (GM) mice or agents such as antibodies or drugs). Using this method, we have robustly screened more than 950 GM mouse lines to identify novel microenvironmental 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

Speak, Anneliese O

Swiatkowska, Agnieszka

Karp, Natasha A

Arends, Mark J

Adams, David J

Van Der Weyden, Louise

Edinburgh Research Explorer

A high-throughput in vivo screening method in the mouse for identifying regulators of metastatic colonization

Crossref

Nature Protocols

https://www.pure.ed.ac.uk/ws/files/46311729/A_high_throughput_in_vivo_screening_method_in_the_mouse_for_identifying_regulators_of_metastatic_colonisation.pdf

A high-throughput in vivo screening method in the mouse for identifying regulators of metastatic colonization

Abstract

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