1. Tension, Free Space, and Cell Damage in a Microfluidic Wound Healing Assay
- Author
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Roger D. Kamm, Paul Matsudaira, Michael P. Murrell, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Mechanical Engineering, Whitehead Institute for Biomedical Research, Kamm, Roger Dale, Murrell, Michael, and Matsudaira, Paul T.
- Subjects
Leading edge ,medicine.medical_specialty ,Materials Science ,Microfluidics ,Cell ,Biophysics ,lcsh:Medicine ,Motility ,Bioengineering ,Biomaterials ,Engineering ,Materials Physics ,Cell Movement ,Cleave ,medicine ,Nanotechnology ,Animals ,Humans ,lcsh:Science ,Biology ,Condensed-Matter Physics ,Cell damage ,Wound Healing ,Multidisciplinary ,Chemistry ,Systems Biology ,Mechanical Engineering ,Physics ,lcsh:R ,Epithelial Cells ,Cell migration ,medicine.disease ,Surgery ,medicine.anatomical_structure ,lcsh:Q ,Biological Assay ,Wound healing ,Research Article ,Biotechnology - Abstract
We use a novel, microfluidics-based technique to deconstruct the classical wound healing scratch assay, decoupling the contribution of free space and cell damage on the migratory dynamics of an epithelial sheet. This method utilizes multiple laminar flows to selectively cleave cells enzymatically, and allows us to present a 'damage free' denudation. We therefore isolate the influence of free space on the onset of sheet migration. First, we observe denudation directly to measure the retraction in the cell sheet that occurs after cell-cell contact is broken, providing direct and quantitative evidence of strong tension within the sheet. We further probe the mechanical integrity of the sheet without denudation, instead using laminar flows to selectively inactivate actomyosin contractility. In both cases, retraction is observed over many cell diameters. We then extend this method and complement the enzymatic denudation with analogies to wounding, including gradients in signals associated with cell damage, such as reactive oxygen species, suspected to play a role in the induction of movement after wounding. These chemical factors are evaluated in combination with the enzymatic cleavage of cells, and are assessed for their influence on the collective migration of a non-abrasively denuded epithelial sheet. We conclude that free space alone is sufficient to induce movement, but this movement is predominantly limited to the leading edge, leaving cells further from the edge less able to move towards the wound. Surprisingly, when coupled with a gradient in ROS to simulate the chemical effects of abrasion however, motility was not restored, but further inhibited., Massachusetts Institute of Technology. Presidential Fellowship, National Institutes of Health (U.S.). Biotechnology Training Fellowship, Singapore-MIT Alliance for Research and Technology, Massachusetts Institute of Biotechnology Training Grant, Massachusetts Institute of Technology (Open-source Funding)
- Published
- 2011
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