Abstract
IntroductionMechanical forces regulate many facets of cell and tissue biology. Studying the effects of forces on cells requires real-time observations of single- and multi-cell dynamics in tissue models during controlled external mechanical input. Many of the existing devices used to conduct these studies are costly and complicated to fabricate, which reduces the availability of these devices to many laboratories.MethodsWe show how to fabricate a simple, low-cost, uniaxial stretching device, with readily available materials and instruments that is compatible with high-resolution time-lapse microscopy of adherent cell monolayers. In addition, we show how to construct a pressure controller that induces a repeatable degree of stretch in monolayers, as well as a custom MATLAB code to quantify individual cell strains.ResultsAs an application note using this device, we show that uniaxial stretch slows down cellular movements in a mammalian epithelial monolayer in a cell density-dependent manner. We demonstrate that the effect on cell movement involves the relocalization of myosin downstream of Rho-associated protein kinase (ROCK).ConclusionsThis mechanical device provides a platform for broader involvement of engineers and biologists in this important area of cell and tissue biology. We used this device to demonstrate the mechanical regulation of collective cell movements in epithelia.
Highlights
Mechanical forces regulate many facets of cell and tissue biology
Using PIVlab, a GUI based particle image velocimetry software, to measure cell migration speeds,35,48,49,52 we show that cellular movement initially slowed down, but the average speed eventually returned to levels observed before the application of stretch
We modeled the mechanical strain of the cell culture membrane of the device under increasing 1 kPa increments of vacuum pressure from 0 to 70 kPa using finite element analysis (FEA) (Fig. 1)
Summary
Mechanical forces regulate many facets of cell and tissue biology. Studying the effects of forces on cells requires real-time observations of single- and multi-cell dynamics in tissue models during controlled external mechanical input. Methods—We show how to fabricate a simple, low-cost, uniaxial stretching device, with readily available materials and instruments that is compatible with high-resolution timelapse microscopy of adherent cell monolayers. Conclusions—This mechanical device provides a platform for broader involvement of engineers and biologists in this important area of cell and tissue biology The fabrication of these devices often requires custom engineering design and access to sophisticated facilities such as a clean room or machine shop, which are not readily available to most laboratories. Many of these devices are incompatible with high-resolution imaging modalities that are indispensable to explore how cells transduce mechanical inputs into downstream signaling outputs. We achieved our goal by using low-cost 3D-printed molds that create optically clear, flexible, silicone-based components, and a vacuumdriven stretching design
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