Abstract
During metastasis, a subset of cancer cells will break away from the primary tumor and invade into the surrounding tissue. Cancer cells which are able to breach the endothelium and enter the blood stream are then transported in the circulation to new target organs where they may seed as a distant metastasis. In order to invade this new organ, the cancer cells must bind to and traverse the vascular wall, a process known as transendothelial migration (TEM) or extravasation. This chapter describes an in vitro approach to automated live cell imaging and analysis of TEM in order to accurately quantify these kinetics and aid the researcher in dissecting the mechanisms of tumor–endothelial interactions during this phase of metastasis.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Reymond N, d’Água BB, Ridley AJ (2013) Crossing the endothelial barrier during metastasis. Nat Rev Cancer 13:858–870
Ley K, Laudanna C, Cybulsky MI, Nourshargh S (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 7:678–689
Follain G, Osmani N, Azevedo AS, Allio G, Mercier L, Karreman MA, Solecki G, Garcia Leòn MJ, Lefebvre O, Fekonja N, Hille C, Chabannes V, Dollé G, Metivet T, Der Hovsepian F, Prudhomme C, Pichot A, Paul N, Carapito R, Bahram S, Ruthensteiner B, Kemmling A, Siemonsen S, Schneider T, Fiehler J, Glatzel M, Winkler F, Schwab Y, Pantel K, Harlepp S, Goetz JG (2018) Hemodynamic forces tune the arrest, adhesion, and extravasation of circulating tumor cells. Dev Cell 45:33–52.e12
Madsen CD, Sahai E (2010) Cancer dissemination—lessons from leukocytes. Dev Cell 19:13–26
Chambers AF, Groom AC, MacDonald IC (2002) Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer 2:563–572
Fidler IJ (1970) Metastasis: quantitative analysis of distribution and fate of tumor emboli labeled with 125 I-5-iodo-2′-deoxyuridine. J Natl Cancer Inst 45:773–782
Mamdouh Z, Mikhailov A, Muller WA (2009) Transcellular migration of leukocytes is mediated by the endothelial lateral border recycling compartment. J Exp Med 206:2795–2808
Hynes WF, Pepona M, Robertson C, Alvarado J, Dubbin K, Triplett M, Adorno JJ, Randles A, Moya ML (2020) Examining metastatic behavior within 3D bioprinted vasculature for the validation of a 3D computational flow model. Sci Adv 6:eabb3308
Offeddu GS, Hajal C, Foley CR, Wan Z, Ibrahim L, Coughlin MF, Kamm RD (2021) The cancer glycocalyx mediates intravascular adhesion and extravasation during metastatic dissemination. Commun Biol 4:255
Coughlin MF, Kamm RD (2020) The use of microfluidic platforms to probe the mechanism of cancer cell extravasation. Adv Healthc Mater 9:e1901410
Rahim S, Üren A (2011) A real-time electrical impedance based technique to measure invasion of endothelial cell monolayer by cancer cells. J Vis Exp 2792
Hilfenhaus G, Mompeón A, Freshman J, Prajapati DP, Hernandez G, Freitas VM, Ma F, Langenbacher AD, Mirkov S, Song D, Cho B-K, Goo YA, Pellegrini M, Chen J-N, Damoiseaux R, Iruela-Arispe ML (2021) A high-content screen identifies drugs that restrict tumor cell extravasation across the endothelial barrier. Cancer Res 81:619–633
Reymond N, Im JH, Garg R, Vega FM, Borda d’Agua B, Riou P, Cox S, Valderrama F, Muschel RJ, Ridley AJ (2012) Cdc42 promotes transendothelial migration of cancer cells through β1 integrin. J Cell Biol 199:653–668
Cailleau R, Olivé M, Cruciger QV (1978) Long-term human breast carcinoma cell lines of metastatic origin: preliminary characterization. In Vitro 14:911–915
Mannion AJ, Odell AF, Taylor A, Jones PF, Cook GP (2019) CD99 regulates cancer cell transendothelial migration and endothelial cell function via CDC42 and actin remodelling. bioRxiv:760934
Acknowledgments
This work was supported by the University of Leeds, including a Faculty of Medicine and Health PhD Scholarship to A.J.M.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Mannion, A.J. (2022). Live Cell Imaging and Analysis of Cancer-Cell Transmigration Through Endothelial Monolayers. In: Benest, A.V. (eds) Angiogenesis. Methods in Molecular Biology, vol 2441. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2059-5_26
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2059-5_26
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2058-8
Online ISBN: 978-1-0716-2059-5
eBook Packages: Springer Protocols