Electrofluidic pressure sensor embedded microfluidic device: a study of endothelial cells under hydrostatic pressure and shear stress combinations.

Various microfluidic cell culture devices have been developed for in vitro cell studies because of their capabilities to reconstitute in vivo microenvironments. However, controlling flows in microfluidic devices is not straightforward due to the wide varieties of fluidic properties of biological samples. Currently, flow observations mainly depend on optical imaging and macro scale transducers, which usually require sophisticated instrumentation and are difficult to scale up. Without real time monitoring, the control of flows can only rely on theoretical calculations and numerical simulations. Consequently, these devices have difficulty in being broadly exploited in biological research. This paper reports a microfluidic device with embedded pressure sensors constructed using electrofluidic circuits, which are electrical circuits built by fluidic channels filled with ionic liquid. A microfluidic device culturing endothelial cells under various shear stress and hydrostatic pressure combinations is developed to demonstrate this concept. The device combines the concepts of electrofluidic circuits for pressure sensing, and an equivalent circuit model to design the cell culture channels. In the experiments, human umbilical vein endothelial cells (HUVECs) are cultured in the device with a continuous medium perfusion, which provides the combinatory mechanical stimulations, while the hydrostatic pressures are monitored in real time to ensure the desired culture conditions. The experimental results demonstrate the importance of real time pressure monitoring, and how both mechanical stimulations affect the HUVEC culture. This developed microfluidic device is simple, robust, and can be easily scaled up for high-throughput experiments. Furthermore, the device provides a practical platform for an in vitro cell culture under well-controlled and dynamic microenvironments.

[1]  Lance L. Munn,et al.  Fluid forces control endothelial sprouting , 2011, Proceedings of the National Academy of Sciences.

[2]  G. Hospers,et al.  Endothelium in vitro: A review of human vascular endothelial cell lines for blood vessel-related research , 2004, Angiogenesis.

[3]  Toshiro Ohashi,et al.  Hydrostatic pressure influences morphology and expression of VE-cadherin of vascular endothelial cells. , 2007, Journal of biomechanics.

[4]  Shuichi Takayama,et al.  Responses of endothelial cells to extremely slow flows. , 2011, Biomicrofluidics.

[5]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[6]  Albert van den Berg Quantitative biosciences from nano to macro Indexed in MEDLINE ! , 2012 .

[7]  Robert M. Nerem,et al.  Oscillatory shear stress and hydrostatic pressure modulate cell-matrix attachment proteins in cultured endothelial cells , 2007, In Vitro Cellular & Developmental Biology - Animal.

[8]  M. Poo,et al.  Endothelial cell polarization and chemotaxis in a microfluidic device. , 2008, Lab on a chip.

[9]  Chien-Chung Peng,et al.  Generation of oxygen gradients in microfluidic devices for cell culture using spatially confined chemical reactions. , 2011, Lab on a chip.

[10]  B. Sumpio,et al.  Increased ambient pressure stimulates proliferation and morphologic changes in cultured endothelial cells , 1994, Journal of cellular physiology.

[11]  Yi-Chung Tung,et al.  Integrated electrofluidic circuits: pressure sensing with analog and digital operation functionalities for microfluidics. , 2012, Lab on a chip.

[12]  R M Nerem,et al.  Endothelial cellular response to altered shear stress. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[13]  M. Ritala,et al.  Integrated photocatalytic micropillar nanoreactor electrospray ionization chip for mimicking phase I metabolic reactions. , 2011, Lab on a chip.

[14]  G. Whitesides,et al.  Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device , 2002, Nature Biotechnology.

[15]  M. Woolkalís,et al.  Regulation of VE-cadherin linkage to the cytoskeleton in endothelial cells exposed to fluid shear stress. , 2002, Experimental cell research.

[16]  Shuichi Takayama,et al.  Individually programmable cell stretching microwell arrays actuated by a Braille display. , 2008, Biomaterials.

[17]  D. Beebe,et al.  Fundamentals of microfluidic cell culture in controlled microenvironments. , 2010, Chemical Society reviews.

[18]  Shuichi Takayama,et al.  Combination of fluid and solid mechanical stresses contribute to cell death and detachment in a microfluidic alveolar model. , 2011, Lab on a chip.

[19]  K. Suh,et al.  A multi-layer microfluidic device for efficient culture and analysis of renal tubular cells. , 2010, Lab on a chip.

[20]  Y. Tung,et al.  Magnet-assisted device-level alignment for the fabrication of membrane-sandwiched polydimethylsiloxane microfluidic devices , 2012 .

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

[22]  B. Mosadegh,et al.  Integrated Elastomeric Components for Autonomous Regulation of Sequential and Oscillatory Flow Switching in Microfluidic Devices , 2010, Nature physics.

[23]  B. Chung,et al.  A microfluidic multi-injector for gradient generation. , 2006, Lab on a chip.

[24]  B L Langille,et al.  Transient and steady-state effects of shear stress on endothelial cell adherens junctions. , 1999, Circulation research.

[25]  Teruo Fujii,et al.  Bile canaliculi formation by aligning rat primary hepatocytes in a microfluidic device. , 2011, Biomicrofluidics.

[26]  L. McIntire,et al.  Mechanical effects on endothelial cell morphology: In vitro assessment , 1986, In Vitro Cellular & Developmental Biology.

[27]  Craig A Simmons,et al.  Macro- and microscale fluid flow systems for endothelial cell biology. , 2010, Lab on a chip.

[28]  Shuichi Takayama,et al.  Computer-controlled microcirculatory support system for endothelial cell culture and shearing. , 2005, Analytical chemistry.

[29]  P. Davies,et al.  Flow-mediated endothelial mechanotransduction. , 1995, Physiological reviews.

[30]  L. McIntire,et al.  Response of cultured endothelial cells to steady flow. , 1984, Microvascular research.

[31]  S Chien,et al.  Shear stress induces spatial reorganization of the endothelial cell cytoskeleton. , 1998, Cell motility and the cytoskeleton.

[32]  B. Sumpio,et al.  Cells in focus: endothelial cell. , 2002, The international journal of biochemistry & cell biology.

[33]  R. Bizios,et al.  Morphological and proliferative responses of endothelial cells to hydrostatic pressure: Role of fibroblast growth factor , 1993, Journal of cellular physiology.

[34]  Shuichi Takayama,et al.  Acoustically detectable cellular-level lung injury induced by fluid mechanical stresses in microfluidic airway systems , 2007, Proceedings of the National Academy of Sciences.

[35]  Donald E Ingber,et al.  Microengineered physiological biomimicry: organs-on-chips. , 2012, Lab on a chip.

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

[37]  Shuichi Takayama,et al.  Computerized microfluidic cell culture using elastomeric channels and Braille displays. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[38]  K. Kinoshita,et al.  Consideration of switching mechanism of binary metal oxide resistive junctions using a thermal reaction model , 2007 .

[39]  R. Franke,et al.  Induction of human vascular endothelial stress fibres by fluid shear stress , 1984, Nature.

[40]  G. Whitesides The origins and the future of microfluidics , 2006, Nature.

[41]  R M Nerem,et al.  Effects of pulsatile flow on cultured vascular endothelial cell morphology. , 1991, Journal of biomechanical engineering.

[42]  Shuichi Takayama,et al.  Microfluidic Endothelium for Studying the Intravascular Adhesion of Metastatic Breast Cancer Cells , 2009, PloS one.

[43]  N. Jeon,et al.  Biological applications of microfluidic gradient devices. , 2010, Integrative biology : quantitative biosciences from nano to macro.

[44]  R. Lal,et al.  Shear stress-induced reorganization of the surface topography of living endothelial cells imaged by atomic force microscopy. , 1994, Circulation research.