Using real-time impedance-based assays to monitor the effects of fibroblast-derived media on the adhesion, proliferation, migration and invasion of colon cancer cells

Abstract

Increasing our knowledge of the mechanisms regulating cell proliferation, migration and invasion are central to understanding tumour progression and metastasis. The local tumour microenvironment contributes to the transformed phenotype in cancer by providing specific environmental cues that alter the cells behaviour and promotes metastasis. Fibroblasts have a strong association with cancer and in recent times there has been some emphasis in designing novel therapeutic strategies that alter fibroblast behaviour in the tumour microenvironment. Fibroblasts produce growth factors, chemokines and many of the proteins laid down in the ECM (extracellular matrix) that promote angiogenesis, inflammation and tumour progression. In this study, we use a label-free RTCA (real-time cell analysis) platform (xCELLigence) to investigate how media derived from human fibroblasts alters cancer cell behaviour. We used a series of complimentary and novel experimental approaches to show HCT116 cells adhere, proliferate and migrate significantly faster in the presence of media from human fibroblasts. As well as this, we used the xCELLigence CIM-plates system to show that HCT116 cells invade matrigel layers aggressively when migrating towards media derived from human fibroblasts. These data strongly suggest that fibroblasts have the ability to increase the migratory and invasive properties of HCT116 cells. This is the first study that provides real-time data on fibroblast-mediated migration and invasion kinetics of colon cancer cells.

Real-time cell analysis of the inhibitory effect of vitamin K2 on adhesion and proliferation of breast cancer cells

“…The RTCA xCELLigence platform used in this study is highly advantageous in comparison to these assays as it facilitates label-free, continuous monitoring of cellular response to compounds [31,32]. It has been shown to correlate very well with the conventional adhesion, viability, migration, and invasion assays and proven to be a highly accurate platform to monitor cell behavior [27,[33][34][35][36][37][38].…”

“…The cells were counted using a hemocytometer. Cells were seeded in each well of the E-plate [27]. The impedance values of each well were automatically monitored by the xCELLigence system and expressed as a cell index (CI) value.…”

“…Cells were carefully washed with water and left overnight to dry. The plates were then read at 590 nmol/L on a spectrophotometer [27].…”

SEM and ECIS Investigation of Cells Cultured on Nanopillar Modified Interdigitated Impedance Electrodes for Analysis of Cell Growth and Cytotoxicity of Potential Anticancer Drugs

“…tissue cultures dishes but they appear more rounded and closer in appearance to what they would look like in a normal in-vivo tissue [30]. The objectives of this research are to study whether the nanopillars allow a successful growth of the cells on them; if, within electrical impedance measurements for cytotoxicity monitoring, it is possible to achieve better sensitivity in the assay compared to the same type of flat electrodes; test different cell lines on the nanostructured electrodes exposed to two different chemicals to determine their cytotoxicity on A549, Balb c3T3 and HeLa cell lines (the chemicals used are an extract of Antrodia Camphorata and Nicotine); imaging the different cell lines immobilized on the flat and nanostructured surfaces to compare their morphology and observe how the surface topography influences the attachment of cell.…”

Microfluidic platform for impedance characterization of endothelial cells under fluid shear stress.

“…They are usually patterned to cover a larger surface area of the substrate surface (~70%) [165], [166] and thus, provide an overall assessment of the entire cell population. IDEs have been used in numerous applications including cell-ECM interactions [167], signal transduction assays [77], cell migration through a cell monolayer [168], barrier function [169] and cell proliferation [170].…”