A tapered channel microfluidic device for comprehensive cell adhesion analysis, using measurements of detachment kinetics and shear stress-dependent motion.

We have developed a method for studying cellular adhesion by using a custom-designed microfluidic device with parallel non-connected tapered channels. The design enables investigation of cellular responses to a large range of shear stress (ratio of 25) with a single input flow-rate. For each shear stress, a large number of cells are analyzed (500-1500 cells), providing statistically relevant data within a single experiment. Besides adhesion strength measurements, the microsystem presented in this paper enables in-depth analysis of cell detachment kinetics by real-time videomicroscopy. It offers the possibility to analyze adhesion-associated processes, such as migration or cell shape change, within the same experiment. To show the versatility of our device, we examined quantitatively cell adhesion by analyzing kinetics, adhesive strength and migration behaviour or cell shape modifications of the unicellular model cell organism Dictyostelium discoideum at 21 °C and of the human breast cancer cell line MDA-MB-231 at 37 °C. For both cell types, we found that the threshold stresses, which are necessary to detach the cells, follow lognormal distributions, and that the detachment process follows first order kinetics. In addition, for particular conditions' cells are found to exhibit similar adhesion threshold stresses, but very different detachment kinetics, revealing the importance of dynamics analysis to fully describe cell adhesion. With its rapid implementation and potential for parallel sample processing, such microsystem offers a highly controllable platform for exploring cell adhesion characteristics in a large set of environmental conditions and cell types, and could have wide applications across cell biology, tissue engineering, and cell screening.

[1]  C W Smith,et al.  Effect of venous shear stress on CD18-mediated neutrophil adhesion to cultured endothelium. , 1990, Blood.

[2]  G. Batchelor,et al.  An Introduction to Fluid Dynamics , 1968 .

[3]  D A Lauffenburger,et al.  Receptor-mediated cell attachment and detachment kinetics. I. Probabilistic model and analysis. , 1990, Biophysical journal.

[4]  G. Whitesides,et al.  Soft lithographic methods for nano-fabrication , 1997 .

[5]  P Ducheyne,et al.  Quantification of cell adhesion using a spinning disc device and application to surface-reactive materials. , 1997, Biomaterials.

[6]  D. Vautier,et al.  Endothelial cell--interactions with polyelectrolyte multilayer films. , 2005, Biomaterials.

[7]  C. Zhu,et al.  Kinetics and mechanics of cell adhesion. , 2000, Journal of biomechanics.

[8]  David A Calderwood,et al.  Forces and Bond Dynamics in Cell Adhesion , 2007, Science.

[9]  David L. Steffen,et al.  The genome of the social amoeba Dictyostelium discoideum , 2005, Nature.

[10]  M. Dembo,et al.  Kinetics of cell detachment: peeling of discrete receptor clusters. , 1994, Biophysical journal.

[11]  Byungkyu Kim,et al.  Label-free, microfluidic separation and enrichment of human breast cancer cells by adhesion difference. , 2007, Lab on a chip.

[12]  P. Lobie,et al.  PAX5α Enhances the Epithelial Behavior of Human Mammary Carcinoma Cells , 2010, Molecular Cancer Research.

[13]  A. L. Koch,et al.  The logarithm in biology. 1. Mechanisms generating the log-normal distribution exactly. , 1966, Journal of theoretical biology.

[14]  E. Elson,et al.  A mechanical function of myosin II in cell motility. , 1995, Journal of cell science.

[15]  Douglas A Lauffenburger,et al.  Microfluidic shear devices for quantitative analysis of cell adhesion. , 2004, Analytical chemistry.

[16]  Alex Groisman,et al.  Quantitative measurements of the strength of adhesion of human neutrophils to a substratum in a microfluidic device. , 2007, Analytical chemistry.

[17]  Y. Bréchet,et al.  Peeling model for cell detachment , 2002, The European physical journal. E, Soft matter.

[18]  P. Pittet,et al.  Fast prototyping using a dry film photoresist: microfabrication of soft-lithography masters for microfluidic structures , 2007 .

[19]  G. Schmid-Schönbein,et al.  Mechanisms for regulation of fluid shear stress response in circulating leukocytes. , 2000, Circulation research.

[20]  A S Rudolph,et al.  Cellular and cytoskeleton morphology and strength of adhesion of cells on self-assembled monolayers of organosilanes. , 1998, Experimental cell research.

[21]  P. Friedl.  Prespecification and plasticity: shifting mechanisms of cell migration. , 2004, Current opinion in cell biology.

[22]  C. Simmons,et al.  Matrix-dependent adhesion of vascular and valvular endothelial cells in microfluidic channels. , 2007, Lab on a chip.

[23]  Peter F. Davies,et al.  Shear Stress Biology of the Endothelium , 2005, Annals of Biomedical Engineering.

[24]  Nitzan Resnick,et al.  Fluid shear stress and the vascular endothelium: for better and for worse. , 2003, Progress in biophysics and molecular biology.

[25]  C. Legrand,et al.  Shear Stress Induced Release of Von Willebrand Factor and Thrombospondin-1 in Uvec Extracellular Matrix Enhances Breast Tumour Cell Adhesion , 2005, Clinical & Experimental Metastasis.

[26]  Y. Bréchet,et al.  Shear flow-induced detachment kinetics of Dictyostelium discoideum cells from solid substrate. , 2002, Biophysical journal.

[27]  D A Lauffenburger,et al.  Maximal migration of human smooth muscle cells on fibronectin and type IV collagen occurs at an intermediate attachment strength , 1993, The Journal of cell biology.

[28]  D. Hammer,et al.  A theoretical analysis for the effect of focal contact formation on cell-substrate attachment strength. , 1993, Biophysical journal.

[29]  James C Baygents,et al.  Detachment of captured cancer cells under flow acceleration in a bio-functionalized microchannel. , 2009, Lab on a chip.

[30]  Douglas A Lauffenburger,et al.  Co-regulation of cell adhesion by nanoscale RGD organization and mechanical stimulus. , 2002, Journal of cell science.

[31]  Thomas Gervais,et al.  Flow-induced deformation of shallow microfluidic channels. , 2006, Lab on a chip.

[32]  Yoshinori Hayakawa,et al.  A Quorum-Sensing Factor in Vegetative Dictyostelium Discoideum Cells Revealed by Quantitative Migration Analysis , 2011, PloS one.

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

[34]  Donald E. Ingber,et al.  The structural and mechanical complexity of cell-growth control , 1999, Nature Cell Biology.

[35]  B. Geiger,et al.  Environmental sensing through focal adhesions , 2009, Nature Reviews Molecular Cell Biology.