Quantitative Analysis of human Cancer Cell Extravasation Using Intravital Imaging

Methods Mol Biol. 2016;1458:27-37

Willetts L, Bond D, Stoletov 1, Lewis JD


Metastasis, or the spread of cancer cells from a primary tumor to distant sites, is the leading cause of cancer-associated death. Metastasis is a complex multi-step process comprised of invasion, intravasation, survival in circulation, extravasation, and formation of metastatic colonies. Currently, in vitro assays are limited in their ability to investigate these intricate processes and do not faithfully reflect metastasis as it occurs in vivo. Traditional in vivo models of metastasis are limited by their ability to visualize the seemingly sporadic behavior of where and when cancer cells spread (Reymond et al., Nat Rev Cancer 13:858-870, 2013). The avian embryo model of metastasis is a powerful platform to study many of the critical steps in the metastatic cascade including the migration, extravasation, and invasion of human cancer cells in vivo (Sung et al., Nat Commun 6:7164, 2015; Leong et al., Cell Rep 8, 1558-1570, 2014; Kain et al., Dev Dyn 243:216-28, 2014; Leong et al., Nat Protoc 5:1406-17, 2010; Zijlstra et al., Cancer Cell 13:221-234, 2008; Palmer et al., J Vis Exp 51:2815, 2011). The chicken chorioallantoic membrane (CAM) is a readily accessible and well-vascularized tissue that surrounds the developing embryo. When the chicken embryo is grown in a shell-less, ex ovo environment, the nearly transparent CAM provides an ideal environment for high-resolution fluorescent microcopy approaches. In this model, the embryonic chicken vasculature and labeled cancer cells can be visualized simultaneously to investigate specific steps in the metastatic cascade including extravasation. When combined with the proper image analysis tools, the ex ovo chicken embryo model offers a cost-effective and high-throughput platform for the quantitative analysis of tumor cell metastasis in a physiologically relevant in vivo setting. Here we discuss detailed procedures to quantify cancer cell extravasation in the shell-less chicken embryo model with advanced fluorescence microscopy techniques.


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Congratulations to first place winner Doug Brown!

Congratulations to PhD grad student Doug Brown for his First Place win at the 2018 University of Alberta Student Falling Walls Lab Competition! Doug’s First Place award also gains him an entry and an all-expenses-paid trip to the highly competitive and prestigious Falling Walls Lab Conference in Berlin, November 8-9, 2018!

Fifteen student presenters pitched their innovative and globally-important research ideas in a short 3 minute presentation to a panel of science and tech-business savvy judges and a large cheering audience on September 19, 2018, at the University of Alberta in a high-stakes “TEDTalk” meets “Dragon’s Den” style. The competition was very strong but Doug gave an inspired speech on breaking down the walls of metastatic cancers using new lipid nanoparticle technology that he is helping to develop during his graduate studies in Dr. John Lewis’ laboratory.

The Sept 19th UofA Falling Walls Lab competition will be featured on Global News on the evening of Sept. 20th, around 6:15 pm. Check it out!

Doug will be taking his pitch to the Falling Walls Lab competition in Berlin, Germany this November where he will compete with many other inspirational students at this international event.

This is doubly exciting for the Lewis lab because John Lewis recently won the UofA Falling Walls Venture competition that was held on Aug 29, 2018 and he will be competing in the Berlin Falling Walls Venture!

Good luck to both Doug and John at the Falling Walls Finale in Berlin!

- Perrin Beatty