Deformable 96-well cell culture plate compatible with high-throughput screening platforms

Abstract

Adherent cells such as endothelial cells sense applied mechanical stretch to adapt to changes in their surrounding mechanical environment. Despite numerous studies, signaling pathways underlying the cellular mechanosensing and adaptation remain to be fully elucidated partly because of the lack of tools that allow for a comprehensive screening approach. Conventionally, multi-well cell culture plates of standard configurations are used for comprehensive analyses in cell biology study to identify key molecules in a high-throughput manner. Given that situation, here we design a 96-well cell culture plate made of elastic silicone and mechanically stretchable using a motorized device. Computational analysis suggested that highly uniform stretch can be applied to each of the wells other than the peripheral wells. Elastic image registration-based experimental evaluation on stretch distributions within individual wells revealed the presence of larger variations among wells compared to those in the computational analysis, but a stretch level of 10%–that has been employed in conventional studies on cellular response to stretch—was almost achieved with our setup. We exposed vascular smooth muscle cells to cyclic stretch using the device to demonstrate morphological repolarization of the cells, i.e. typical cellular response to cyclic stretch. Because the deformable multi-well plate validated here is compatible with other high-throughput screening-oriented technologies, we expect this novel system to be utilized for future comprehensive analyses of stretch-related signaling pathways.

The Form and Function of PIEZO2

“…Membrane stretch, pull, indentation, fluid shear stress, and substrate deformation are examples of stimuli that have been classically applied in a low-throughput setting (30,42,66,165,166). However, some of these techniques are now being scaled up and may be amenable for mechanical screening of PIEZO2 (167)(168)(169)(170)(171)(172).…”

Multivariate landscapes constructed by Bayesian estimation over five hundred microbial electrochemical time profiles

“…While the maximum I c has the lowest deviation percentage from the mean, the other two also lie within the SD, which is comparable with established high-throughput systems. 31 , 32 Thus, based on the data variation, our system demonstrated high reproducibility through all wells in the 96-well plate.…”

The role of mechanobiology in bone and cartilage model systems in characterizing initiation and progression of osteoarthritis

“…However, ongoing research is significantly increasing the throughput of these types of stretching systems. 203,204 Aside from cell stretching, fluid flow systems allow the investigation of the effects of shear on osteochondral cells. These typically involve application of controlled unidirectional or oscillatory fluid movement in a cell culture chamber and range from rocking cultures and cone viscometers to complex microfluidic devices.…”