Controlled electromechanical cell stimulation on-a-chip

Stem cell research has yielded promising advances in regenerative medicine, but standard assays generally lack the ability to combine different cell stimulations with rapid sample processing and precise fluid control. In this work, we describe the design and fabrication of a micro-scale cell stimulator capable of simultaneously providing mechanical, electrical, and biochemical stimulation, and subsequently extracting detailed morphological and gene-expression analysis on the cellular response. This micro-device offers the opportunity to overcome previous limitations and recreate critical elements of the in vivo microenvironment in order to investigate cellular responses to three different stimulations. The platform was validated in experiments using human bone marrow mesenchymal stem cells. These experiments demonstrated the ability for inducing changes in cell morphology, cytoskeletal fiber orientation and changes in gene expression under physiological stimuli. This novel bioengineering approach can be readily applied to various studies, especially in the fields of stem cell biology and regenerative medicine.

[1]  Min Zhao,et al.  Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-gamma and PTEN. , 2006, Nature.

[2]  Y. Morsi,et al.  From mechanical stimulation to biological pathways in the regulation of stem cell fate , 2014, Cell biochemistry and function.

[3]  Milica Radisic,et al.  Functional assembly of engineered myocardium by electrical stimulation of cardiac myocytes cultured on scaffolds , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Payam Akhyari,et al.  A novel miniaturized multimodal bioreactor for continuous in situ assessment of bioartificial cardiac tissue during stimulation and maturation. , 2011, Tissue engineering. Part C, Methods.

[5]  R. Kelly,et al.  Continual electric field stimulation preserves contractile function of adult ventricular myocytes in primary culture. , 1994, The American journal of physiology.

[6]  S Chien,et al.  Effects of disturbed flow on endothelial cells. , 1998, Journal of biomechanical engineering.

[7]  Stefan Wagner,et al.  Murine and human pluripotent stem cell-derived cardiac bodies form contractile myocardial tissue in vitro. , 2013, European heart journal.

[8]  H. Drexler,et al.  Clinical applications of stem cells for the heart. , 2005, Circulation research.

[9]  Robert F Service Bioengineering. Lung-on-a-chip breathes new life into drug discovery. , 2012, Science.

[10]  Evelyn K F Yim,et al.  Nanotopography/mechanical induction of stem-cell differentiation. , 2010, Methods in cell biology.

[11]  Sottile,et al.  Multi-lineage potential of human mesenchymal stem cells following clonal expansion. , 2001, Journal of musculoskeletal & neuronal interactions.

[12]  Milica Radisic,et al.  Electrical stimulation systems for cardiac tissue engineering , 2009, Nature Protocols.

[13]  W. Giles,et al.  Monophasic Versus Biphasic Cardiac Stimulation: Mechanism of Decreased Energy Requirements , 1990, Pacing and clinical electrophysiology : PACE.

[14]  Milica Radisic,et al.  Challenges in cardiac tissue engineering. , 2010, Tissue engineering. Part B, Reviews.

[15]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[16]  G. Forte,et al.  Cooperation of Biological and Mechanical Signals in Cardiac Progenitor Cell Differentiation , 2011, Advanced materials.

[17]  D. Ingber,et al.  Microfluidic organs-on-chips , 2014, Nature Biotechnology.

[18]  W. Blau,et al.  The electrical stimulation of carbon nanotubes to provide a cardiomimetic cue to MSCs. , 2012, Biomaterials.

[19]  Leslie Tung,et al.  Electrical pacing counteracts intrinsic shortening of action potential duration of neonatal rat ventricular cells in culture. , 2006, Journal of molecular and cellular cardiology.

[20]  Ya-Pu Zhao,et al.  Deformation of PDMS membrane and microcantilever by a water droplet: comparison between Mooney-Rivlin and linear elastic constitutive models. , 2009, Journal of colloid and interface science.

[21]  Jianwen Luo,et al.  Biomimetic perfusion and electrical stimulation applied in concert improved the assembly of engineered cardiac tissue , 2012, Journal of tissue engineering and regenerative medicine.

[22]  Kerstin Pingel,et al.  50 Years of Image Analysis , 2012 .

[23]  Joel Voldman,et al.  Fluid shear stress primes mouse embryonic stem cells for differentiation in a self‐renewing environment via heparan sulfate proteoglycans transduction , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  Milica Radisic,et al.  Interactive effects of surface topography and pulsatile electrical field stimulation on orientation and elongation of fibroblasts and cardiomyocytes. , 2007, Biomaterials.

[25]  D. Ingber,et al.  Reconstituting Organ-Level Lung Functions on a Chip , 2010, Science.

[26]  Hanry Yu,et al.  Stem cells in microfluidics , 2009, Biotechnology progress.

[27]  Gianfranco Beniamino Fiore,et al.  Monophasic and biphasic electrical stimulation induces a precardiac differentiation in progenitor cells isolated from human heart. , 2014, Stem cells and development.

[28]  W. Claycomb,et al.  Effect of mechanical loading on three-dimensional cultures of embryonic stem cell-derived cardiomyocytes. , 2008, Tissue engineering. Part A.

[29]  Tal Dvir,et al.  Electric field stimulation integrated into perfusion bioreactor for cardiac tissue engineering. , 2010, Tissue engineering. Part C, Methods.

[30]  S. Ferrari,et al.  Author contributions , 2021 .

[31]  Yubo Fan,et al.  Endothelium oriented differentiation of bone marrow mesenchymal stem cells under chemical and mechanical stimulations. , 2010, Journal of biomechanics.

[32]  N. Tran,et al.  Effect of uniaxial stretching on rat bone mesenchymal stem cell: Orientation and expressions of collagen types I and III and tenascin‐C , 2008, Cell biology international.

[33]  Andrea Pavesi,et al.  Microfabrication and microfluidics for muscle tissue models. , 2014, Progress in biophysics and molecular biology.

[34]  Jennifer S. Park,et al.  Differential effects of equiaxial and uniaxial strain on mesenchymal stem cells , 2004, Biotechnology and bioengineering.

[35]  A. Redaelli,et al.  Electrical conditioning of adipose‐derived stem cells in a multi‐chamber culture platform , 2014, Biotechnology and bioengineering.

[36]  Marco Rasponi,et al.  How to embed three-dimensional flexible electrodes in microfluidic devices for cell culture applications. , 2011, Lab on a chip.

[37]  Chelsey S Simmons,et al.  Microsystems for biomimetic stimulation of cardiac cells. , 2012, Lab on a chip.

[38]  N. Tandon,et al.  Optimization of electrical stimulation parameters for cardiac tissue engineering , 2011, Journal of tissue engineering and regenerative medicine.

[39]  Milica Radisic,et al.  Biphasic electrical field stimulation aids in tissue engineering of multicell-type cardiac organoids. , 2011, Tissue engineering. Part A.

[40]  Dimitrios Vavylonis,et al.  Interactive, computer-assisted tracking of speckle trajectories in fluorescence microscopy: application to actin polymerization and membrane fusion. , 2011, Biophysical journal.

[41]  Nam-Trung Nguyen,et al.  A polymeric cell stretching device for real-time imaging with optical microscopy , 2013, Biomedical Microdevices.

[42]  Donald E Ingber,et al.  Microfabrication of human organs-on-chips , 2013, Nature Protocols.

[43]  Nitish Thakor,et al.  Valve-based microfluidic compression platform: single axon injury and regrowth. , 2011, Lab on a chip.

[44]  Min Zhao,et al.  Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-γ and PTEN , 2006, Nature.

[45]  Swathi Rangarajan,et al.  Use of Flow, Electrical, and Mechanical Stimulation to Promote Engineering of Striated Muscles , 2013, Annals of Biomedical Engineering.

[46]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[47]  Ebrahim Ghafar-Zadeh,et al.  Engineered approaches to the stem cell microenvironment for cardiac tissue regeneration. , 2011, Lab on a chip.

[48]  Quanfang Chen,et al.  Thickness-dependent mechanical properties of polydimethylsiloxane membranes , 2009 .

[49]  S. Chien Effects of Disturbed Flow on Endothelial Cells , 2008, Annals of Biomedical Engineering.

[50]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[51]  Zhonggang Feng,et al.  An electro-tensile bioreactor for 3-D culturing of cardiomyocytes , 2005, IEEE Engineering in Medicine and Biology Magazine.

[52]  N. Bursac,et al.  Effect of Electromechanical Stimulation on the Maturation of Myotubes on Aligned Electrospun Fibers , 2008, Cellular and molecular bioengineering.

[53]  Yu Sun,et al.  Microfabricated arrays for high-throughput screening of cellular response to cyclic substrate deformation. , 2010, Lab on a chip.

[54]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.