An Automated High-Content Assay for Tumor Cell Migration through 3-Dimensional Matrices

Abstract

High-content tumor cell migration assays in 3-dimensional (3D) extracellular matrix are a powerful tool for modeling and understanding the biology of this critical step in the process of metastasis. Currently available methods offer very limited throughput and are not amenable to studies of comparative pharmacology or small-scale screening. The authors present an automated approach to high-content tumor cell migration assays. A standard screening-sized plate with an array of microchannels was designed and constructed from common thermoplastics. After filling the channels with 3D matrix, cells were placed at one end of the channel, and migration into the channel was monitored via an imaging system. All liquid-handling steps were performed by standard liquid-handling robotics. Tumor cell migration in the channel was truly 3D and correlated with metastatic potential. The information-rich data from these assays were used to rank the potency of compounds inhibiting migration through 3D collagen as well as to gain additional insights into the compounds' activities related to cell health. This approach is compatible with a variety of multiparametric, morphological, and/or kinetic readouts.

Cell Migration and Invasion Assays as Tools for Drug Discovery

“…An example schematic is provided in Figure 3. Specifically, a 192 individually addressable microchannel array device was developed at BellBrook Labs which uses droplet-based passive pumping to seed cells, exchange media, and then fix and stain cells [39,40]. This device can be fully automated in conjunction with liquid handling and microscopic imaging instruments [40].…”

“…A microfluidic assay was used to demonstrate tumor cell migration through 3-dimensional matrices [40]. In this assay, the microchannels were filled with either a rat tail collagen I or Matrigel™ which was allowed to gel prior to the addition of 2,000 PC-3M cells in a 2.5 μL droplet.…”

“…Images were captured using a microscope with an automated stage and camera in conjunction with Metamorph® software for image analysis and cell counts. A Z′ factor of 0.44 was calculated for this assay as determined by treatment of cells with 5 μM blebbistatin as a pre-invasion control compared with untreated controls permitted to invade in the absence of compounds [40]. …”

Quantitative analysis of gradient sensing: towards building predictive models of chemotaxis in cancer

“…Another, perhaps more generally reliable and accessible, approach to obtain information-rich, single-cell, spatially relevant data for relational models, is the employment of high-content imaging [58]. Two examples illustrate the usefulness of this approach for studying directed migration.…”

A High Content Drug Screen Identifies Ursolic Acid as an Inhibitor of Amyloid β Protein Interactions with Its Receptor CD36

“…The images can subsequently be analyzed by imaging software to provide very detailed information about the cells in question. The technology has been used previously to find small molecule regulators of G protein-coupled receptors (14), develop assays to find inhibitors of metastasis (15), and to find compounds that increase the growth of pancreatic ␤ cells (16). Using this approach we developed an assay to quantify binding of A␤ to CHO cells stably transfected with CD36.…”