Abstract

BackgroundCell invasion through extracellular matrix (ECM) is a critical step in tumor metastasis. To study cell invasion in vitro, the internal microenvironment can be simulated via the application of 3D models.ResultsThis study presents a method for 3D invasion examination using microcarrier-based spheroids. Cell invasiveness can be evaluated by quantifying cell dispersion in matrices or tracking cell movement through time-lapse imaging. It allows measuring of cell invasion and monitoring of dynamic cell behavior in three dimensions. Here we show different invasive capacities of several cell types using this method. The content and concentration of matrices can influence cell invasion, which should be optimized before large scale experiments. We also introduce further analysis methods of this 3D invasion assay, including manual measurements and homemade semi-automatic quantification. Finally, our results indicate that the position of spheroids in a matrix has a strong impact on cell moving paths, which may be easily overlooked by researchers and may generate false invasion results.ConclusionsIn all, the microcarrier-based spheroids 3D model allows exploration of adherent cell invasion in a fast and highly reproducible way, and provides informative results on dynamic cell behavior in vitro.

Highlights

  • Cell invasion through extracellular matrix (ECM) is a critical step in tumor metastasis

  • Among them the Transwell invasion assay, or Boyden chamber assay, is widely used. It includes seeding cells on a layer of ECM gel pre-coated on top of a porous membrane, and assessing cell invasion by measuring the number of cells passing through the ECM gel

  • In this study we present a method for Three dimensions (3D) invasion examination and introduce various measurements according to different experimental settings and requirements

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Summary

Introduction

Cell invasion through extracellular matrix (ECM) is a critical step in tumor metastasis. Malignant tumors have the potential to metastasize from the original tissue to a distant site, which is the main cause of morbidity and mortality in tumor patients During this process, the basic but decisive step is migration of tumor cells through the extracellular matrix (ECM) either towards lymph and blood vessels, or to a secondary site after survival in circulation [1]. There are a diversity of strategies for cells movement, either individually (e.g. amoeboid or mesenchymal migration) or collectively (multicellular streaming, cluster, strand or sheet), which are based on cell-cell adhesion and cell-matrix interaction [3,4,5] This activity can be simulated and observed by. The chamber invasion assay is straightforward to quantify invading cells induced by chemoattractants [6] or internal gene regulation [7]. We describe a microcarrierbased spheroid model to investigate dynamic cell behavior in three dimensional matrices

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