In vitro Cell Migration, Invasion, and Adhesion Assays: From Cell Imaging to Data Analysis.

Jordi Pijuan,Carla Barceló,Anaïs Panosa,Anna Macià,Oscar Maiques,Pol Sisó,David F Moreno,Rosa M Marti

doi:10.3389/fcell.2019.00107

Abstract

Cell migration is a key procedure involved in many biological processes including embryological development, tissue formation, immune defense or inflammation, and cancer progression. How physical, chemical, and molecular aspects can affect cell motility is a challenge to understand migratory cells behavior. In vitro assays are excellent approaches to extrapolate to in vivo situations and study live cells behavior. Here we present four in vitro protocols that describe step-by-step cell migration, invasion and adhesion strategies and their corresponding image data quantification. These current protocols are based on two-dimensional wound healing assays (comparing traditional pipette tip-scratch assay vs. culture insert assay), 2D individual cell-tracking experiments by live cell imaging and three-dimensional spreading and transwell assays. All together, they cover different phenotypes and hallmarks of cell motility and adhesion, providing orthogonal information that can be used either individually or collectively in many different experimental setups. These optimized protocols will facilitate physiological and cellular characterization of these processes, which may be used for fast screening of specific therapeutic cancer drugs for migratory function, novel strategies in cancer diagnosis, and for assaying new molecules involved in adhesion and invasion metastatic properties of cancer cells.

Highlights

Cell migration is a crucial process where cells must be able to change and reach their proper position in a given environment to execute their function
We studied the effect of Ttype calcium channel (TTCC) blocker (Mibefradil) in melanoma individual cell migration
When there is rich media in the upper and lower chamber, there is not significant decrease in migration by Mibefradil treatment (p = 0.6358) because there is no chemoattractant gradient to favor migration directionality (Figure 4D). All these results suggest that Fetal bovine serum (FBS) chemoattractant increases cell migration capacity, but this effect can be attenuated with Mibefradil treatment, which reduces migration rate by blocking the autophagic flux in M3 melanoma cells (Macià et al, 2015; Maiques et al, 2018)

Summary

Introduction

Cell migration is a crucial process where cells must be able to change and reach their proper position in a given environment to execute their function (te Boekhorst et al, 2016). In multicellular organisms, this phenomenon plays an important role in gastrulation, embryonic morphogenesis, development of the nervous system, tissue homeostasis, and immune cell trafficking. In cancer development and progression, invasion, and metastasis occurs when tumor cells disseminate from the primary tumor spreading through the circulatory and lymphatic systems, invade across the basement membranes and endothelial walls and colonize distant organs. Cancer cells movement to peripheral organs, and their resultant destruction, constitutes a primary cause of cancer-associated morbidity and mortality (Xu et al, 2018)

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