Matsui TS, Ito K, Kaunas R, Sato M, Deguchi S. Actin stress fibers are at a tipping point between conventional shortening and rapid disassembly at physiological levels of MgATP. Biochem Biophys Res Commun. 2010;395: 301–306. pmid:20353757 Cells sense mechanical stretch to regulate their morphology and function [ 1]. Cellular response to stretch has been demonstrated to play a pivotal role in developmental process [ 2, 3] as well as in health maintenance [ 4]. One example includes the behavior of endothelial cells that orients their morphological polarization into a direction perpendicular to the direction of stretch upon heartbeat-induced cyclic stretch [ 5– 8]. This endothelial cellular response to orient away from stretch has been implicated in suppression of atherosclerosis to circumvent prolonged activation of pro-inflammatory signals [ 5, 9, 10].

Hirata H, Tatsumi H, Sokabe M. Mechanical forces facilitate actin polymerization at focal adhesions in a zyxin-dependent manner. J Cell Sci. 2008;121: 2795–2804. pmid:18682496

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J. G. Rodrigues, M. Balmana, J. A. Macedo, J. Pocas, A. Fernandes, J. C. M. de-Freitas-Junior, S. S. Pinho, J. Gomes, A. Magalhaes, C. Gomes, S. Mereiter and C. A. Reis, Cell. Immunol., 2018, 333, 46–57 CrossRef CAS PubMed.

Kamble H, Barton MJ, Jun M, Park S, Nguyen N-T. Cell stretching devices as research tools: engineering and biological considerations. Lab Chip. Royal Society of Chemistry; 2016;16: 3193–3203. pmid:27440436 Here, we developed the deformable 96-well cell culture plate with the future perspective of coupling with other 96-well-based technologies such as siRNA/shRNA/drug libraries, automated fluorescence plate readers, and micro-channel pipettes. Given its universal design, we expect our system to be utilized, efficiently together with such existing technologies, as a platform allowing for comprehensive analyses of cellular mechanobiological processes. Corning® 96-well Clear V-Bottom Polystyrene Not Treated Microplate, 25 per Bag, without Lid, Nonsterile Corning® 96-well EIA/RIA Easy Wash™ Clear Flat Bottom Polystyrene Not Treated Microplate, 25 per Bag, without Lid, Nonsterile Abiko H, Fujiwara S, Ohashi K, Hiatari R, Mashiko T, Sakamoto N, et al. Rho guanine nucleotide exchange factors involved in cyclic-stretch-induced reorientation of vascular endothelial cells. J Cell Sci. 2015;128: 1683–1695. pmid:25795300

H. Hinneburg, P. Korac, F. Schirmeister, S. Gasparov, P. H. Seeberger, V. Zoldos and D. Kolarich, Mol. Cell. Proteomics, 2017, 16, 524–536 CrossRef CAS PubMed. Corning® 96-well Clear Flat Bottom Polystyrene TC-treated Microplates, Individually Wrapped, with Lid, Sterile Cellular response to stretch has been extensively studied, but conventional experimental systems are often constituted from single or only 6 wells partly because the majority of the previous studies focused on imaging of the dynamics of specific individual cells/molecules [ 13, 18, 32– 37] or examining changes in mRNA expression [ 22, 38] rather than performing an assay for molecular screening, the last of which generally requires huge numbers of screening trials. For example, a screening study was recently reported to reveal the Rho-GEFs responsible for the cyclic stretch-induced repolarization from 63 candidate molecules [ 6]. Here, they employed a stretch chamber with a single well of a 20 x 20 mm 2 cell culture area and a Rho-GEFs-targeted shRNA library, which we guess might took long time to complete and was costly to prepare large amounts of reagents. In this regard, our strategy of combining the stretch chamber and library directly at the small individual well levels can highly improve the screening throughput and cost. In addition to such candidate gene screenings, our system is useful as well in compound screening with various reagent libraries to suppress or rescue cellular responses altered by gene mutations. To evaluate the actual strain of the plate in response to motions of the motors connected via several mechanical links, the entire plate was imaged during cyclic stretching ( Fig 2B). Quantification of the captured sequential images indicated that indeed the entire plate exhibited a desired sinusoidal 7% stretch pattern as consistent with the programed motions of the motors ( Fig 2C).