Individually programmable cell stretching microwell arrays actuated by a Braille display.

Cell culture systems are often static and are therefore nonphysiological. In vivo, many cells are exposed to dynamic surroundings that stimulate cellular responses in a process known as mechanotransduction. To recreate this environment, stretchable cell culture substrate systems have been developed, however, these systems are limited by being macroscopic and low throughput. We have developed a device consisting of 24 miniature cell stretching chambers with flexible bottom membranes that are deformed using the computer-controlled, piezoelectrically actuated pins of a Braille display. We have also developed efficient image capture and analysis protocols to quantify morphological responses of the cells to applied strain. Human dermal microvascular endothelial cells (HDMECs) were found to show increasing degrees of alignment and elongation perpendicular to the radial strain in response to cyclic stretch at increasing frequencies of 0.2, 1, and 5 Hz, after 2, 4, and 12h. Mouse myogenic C2C12 cells were also found to align in response to the stretch, while A549 human lung adenocarcinoma epithelial cells did not respond to stretch.

[1]  Konstantin G. Birukov,et al.  Stretch affects phenotype and proliferation of vascular smooth muscle cells , 1995, Molecular and Cellular Biochemistry.

[2]  A. Folch,et al.  Large-scale single-cell trapping and imaging using microwell arrays. , 2005, Analytical chemistry.

[3]  Thomas K. Borg,et al.  Effects of cyclic mechanical stimulation of the cellular components of the heart: In vitro , 2007, In Vitro Cellular & Developmental Biology.

[4]  P. Dartsch,et al.  Response of cultured endothelial cells to mechanical stimulation , 1989, Basic Research in Cardiology.

[5]  Herman H. Vandenburgh,et al.  A computerized mechanical cell stimulator for tissue culture: Effects on skeletal muscle organogenesis , 1988, In Vitro Cellular & Developmental Biology.

[6]  Ali Khademhosseini,et al.  A microwell array system for stem cell culture. , 2008, Biomaterials.

[7]  L V McIntire,et al.  Cyclical strain effects on production of vasoactive materials in cultured endothelial cells , 1992, Journal of cellular physiology.

[8]  K. Badr,et al.  Continuous stretch-relaxation in culture alters rat mesangial cell morphology, growth characteristics, and metabolic activity. , 1992, Laboratory investigation; a journal of technical methods and pathology.

[9]  A. Banes,et al.  A new vacuum-operated stress-providing instrument that applies static or variable duration cyclic tension or compression to cells in vitro. , 1985, Journal of cell science.

[10]  K. Audus,et al.  Characterization of the A549 cell line as a type II pulmonary epithelial cell model for drug metabolism. , 1998, Experimental cell research.

[11]  H. Wirtz,et al.  Calcium mobilization and exocytosis after one mechanical stretch of lung epithelial cells. , 1990, Science.

[12]  Nico Westerhof,et al.  Effect of cyclic axial stretch of rat arteries on endothelial cytoskeletal morphology and vascular reactivity. , 2003, Journal of biomechanics.

[13]  K. Esser,et al.  Intracellular signaling specificity in response to uniaxial vs. multiaxial stretch: implications for mechanotransduction. , 2005, American journal of physiology. Cell physiology.

[14]  B. Sumpio,et al.  Strain activation of bovine aortic smooth muscle cell proliferation and alignment: Study of strain dependency and the role of protein kinase A and C signaling pathways , 1997, Journal of cellular physiology.

[15]  Shuichi Takayama,et al.  Handheld recirculation system and customized media for microfluidic cell culture. , 2006, Lab on a chip.

[16]  J. H. Chen,et al.  Strain fields on cell stressing devices employing clamped circular elastic diaphragms as substrates. , 1992, Journal of biomechanical engineering.

[17]  T. Takemasa,et al.  Amplitude-dependent stress fiber reorientation in early response to cyclic strain. , 1997, Experimental cell research.

[18]  G. Whitesides,et al.  Patterning proteins and cells using soft lithography. , 1999, Biomaterials.

[19]  P. Standley,et al.  Cyclic stretch induces vascular smooth muscle cell alignment via NO signaling. , 2002, American journal of physiology. Heart and circulatory physiology.

[20]  D. Tschumperlin,et al.  Deformation-induced injury of alveolar epithelial cells. Effect of frequency, duration, and amplitude. , 2000, American journal of respiratory and critical care medicine.

[21]  D. Brunette,et al.  Mechanical stretching increases the number of cultured bone cells synthesizing DNA and alters their pattern of protein synthesis , 1985, Calcified Tissue International.

[22]  P. Standley,et al.  Erratum: Cyclic stretch induces vascular smooth muscle cell alignment via NO signaling (American Journal of Physiology - Heart and Circulatory Physiology (November 2002) 283 (H1907-H1914)) , 2002 .

[23]  A. J. Reid,et al.  Endothelial cell alignment on cyclically-stretched silicone surfaces , 2004, Journal of materials science. Materials in medicine.

[24]  B. Sumpio,et al.  Effect of cyclic stretch on endothelial cells from different vascular beds. , 1991, Circulatory shock.

[25]  S. Ye,et al.  Magnitude-dependent regulation of pulmonary endothelial cell barrier function by cyclic stretch. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[26]  Milan Mrksich,et al.  Electrochemical desorption of self-assembled monolayers noninvasively releases patterned cells from geometrical confinements. , 2003, Journal of the American Chemical Society.

[27]  P. Dobrin,et al.  Mechanical properties of arteries , 1978, Physiological reviews.

[28]  Takako Yamada,et al.  Involvement of SA channels in orienting response of cultured endothelial cells to cyclic stretch. , 1998, American journal of physiology. Heart and circulatory physiology.

[29]  R. Zare,et al.  Construction of microfluidic chips using polydimethylsiloxane for adhesive bonding. , 2005, Lab on a chip.

[30]  William E. Kraus,et al.  Orientation and length of mammalian skeletal myocytes in response to a unidirectional stretch , 2000, Cell and Tissue Research.

[31]  J. Frangos,et al.  Strain Rate Mechanotransduction in Aligned Human Vascular Smooth Muscle Cells , 2003, Annals of Biomedical Engineering.

[32]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[33]  E K Fram,et al.  Morphometric characteristics of cells in the alveolar region of mammalian lungs. , 2015, The American review of respiratory disease.

[34]  J. R. Scotti,et al.  Available From , 1973 .

[35]  Sean P. Palecek,et al.  3-D microwell culture of human embryonic stem cells. , 2006, Biomaterials.

[36]  S. Pollack,et al.  The proliferative and synthetic response of isolated calvarial bone cells of rats to cyclic biaxial mechanical strain. , 1991, The Journal of bone and joint surgery. American volume.

[37]  T. Ushida,et al.  Mechanical stretch is a down-regulatory signal for differentiation of C2C12 myogenic cells , 2001 .

[38]  G. N. Antonova,et al.  Mechano-chemical control of human endothelium orientation and size , 1989, The Journal of cell biology.

[39]  S. Gorfien,et al.  Effects of biaxial deformation on pulmonary artery endothelial cells , 1989, Journal of cellular physiology.

[40]  R. Adam,et al.  Mechanical Strain Delivers Anti-apoptotic and Proliferative Signals to Gingival Fibroblasts , 2004, Journal of dental research.

[41]  R. Buck Behavior of vascular smooth muscle cells during repeated stretching of the substratum in vitro. , 1983, Atherosclerosis.

[42]  S. Chien,et al.  Effects of mechanical forces on signal transduction and gene expression in endothelial cells. , 1998, Hypertension.

[43]  P. Weinhold,et al.  Strain profiles for circular cell culture plates containing flexible surfaces employed to mechanically deform cells in vitro. , 1994, Journal of biomechanics.

[44]  Thomas J. Burkholder,et al.  Uniaxial strain system to investigate strain rate regulation in vitro , 2001 .