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. 2011 Mar 11;3(1):107-24.
doi: 10.3390/pharmaceutics3010107.

Cell migration and invasion assays as tools for drug discovery

Keren I Hulkower  1 , Renee L Herber
Affiliations

Cell migration and invasion assays as tools for drug discovery

Keren I Hulkower et al. Pharmaceutics. .

Abstract

Cell migration and invasion are processes that offer rich targets for intervention in key physiologic and pathologic phenomena such as wound healing and cancer metastasis. With the advent of high-throughput and high content imaging systems, there has been a movement towards the use of physiologically relevant cell-based assays earlier in the testing paradigm. This allows more effective identification of lead compounds and recognition of undesirable effects sooner in the drug discovery screening process. This article will review the effective use of several principle formats for studying cell motility: scratch assays, transmembrane assays, microfluidic devices and cell exclusion zone assays.

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Figures

Figure 1.
Figure 1.
Scratch Assay. A wound is introduced into a confluent monolayer of cells (A) by drawing a tip across the cell layer (B). The denuded area is imaged to measure the boundary of the wound at pre-migration (C) and after cells have migrated inward to fill the void (D).
Figure 2.
Figure 2.
Transmembrane/Boyden Chamber Assay. A membrane insert is used to establish 2 compartments in a well (A). Cells are added to the upper compartment (B) and migrate through the membrane (C). Cell migration is measured by counting the number of cells on the underside of the membrane (D). Assay options include coating the membrane with a matrix protein and adding a chemoattractant to the lower compartment.
Figure 3.
Figure 3.
Microfluidic Assay. The device provides 2 ports for reagent delivery (A). Cells alone or in a matrix are introduced into the smaller port and adhere to the chamber bottom (B). Test agents are added to the larger port and a gradient sets up based on surface tension (C). Cells can be imaged to measure migration in response to the test agent (D).
Figure 4.
Figure 4.
Cell Exclusion Zone Assay. Cells are seeded around a barrier (A) and adhere to the well bottom. The barrier is removed (B) to reveal a void available for cell movement. The cells are imaged at pre-migration (C) and after cells have migrated inward to fill the void (D). Assay options include coating the assay well with a matrix and adding an overlay of matrix to create a 3-dimensional assay.
Figure 5.
Figure 5.
Comparison of MDA-MB-231 human breast epithelial cell migration using the Oris™ Cell Migration Assay and the scratch assay. MDA-MB-231 cells were seeded into collagen I coated Oris™ assay plates with stoppers at 25,000 cells/ 100 μL or in collagen I coated 6-well plates at 500,000 cells/2 mL. Once confluent monolayers were formed, the cells were serum starved for 24 hours. To initiate migration, the stoppers were removed from the Oris™ assay plate and the monolayers were scratched using a 1,000 μL pipet tip in the 6-well plates. The media in both assays was replaced with serum-containing media. Representative phase images of pre-migration (A and C) and migration after 20 hours (B and D) in the Oris™ (A and B) and the scratch (C and D) assay were captured using a Zeiss Axiovert microscope with an attached CCD camera. Scale bar = 500 μm. Graph of 4 independent experiments comparing cell migration using the Oris™ and scratch assays in parallel (E). Images were analyzed using ImageJ analysis software and data presented as average percent closure ± SD (n ≥ 12 replicates).
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