Using a co-culture microsystem for cell migration under fluid shear stress.

We have successfully developed a microsystem to co-cultivate two types of cells with a minimum defined gap of 50 μm, and to quantitatively study the impact of fluid shear stress on the mutual influence of cell migration velocity and distance. We used the hydrostatic pressure to seed two different cells, endothelial cells (ECs) and smooth muscle cells (SMCs), on opposite sides of various gap sizes (500 μm, 200 μm, 100 μm, and 50 μm). After cultivating the cells for 12 h and peeling the co-culture microchip from the culture dish, we studied the impacts of gap size on the migration of either cell type in the absence or presence of fluid shear stress (7 dyne cm(-2) and 12 dyne cm(-2)) influence. We found that both gap size and shear stress have profound influence on cell migration. Smaller gap sizes (100 μm and 50 μm) significantly enhanced cell migration, suggesting a requirement of an effective concentration of released factor(s) by either cell type in the gap region. Flow-induced shear stress delayed the migration onset of either cell type in a dose-dependent manner regardless of the gap size. Moreover, shear stress-induced decrease of cell migration becomes evident when the gap size was 500 μm. We have developed a co-culture microsystem for two kinds of cells and overcome the conventional difficulties in observation and mixed culture, and it would have more application for bio-manipulation and tissue repair engineering.

[1]  David L. Allara,et al.  Critical issues in applications of self-assembled monolayers , 1995 .

[2]  P Connolly,et al.  Cell guidance by micropatterned adhesiveness in vitro. , 1992, Journal of cell science.

[3]  S. Carter,et al.  Principles of Cell Motility: The Direction of Cell Movement and Cancer Invasion , 1965, Nature.

[4]  Keiji Naruse,et al.  Analysis of cyclic-stretching responses using cell-adhesion-patterned cells. , 2008, Journal of biotechnology.

[5]  M. Toner,et al.  Microengineering of cellular interactions. , 2000, Annual review of biomedical engineering.

[6]  G. Whitesides,et al.  Topographical Micropatterning of Poly(dimethylsiloxane) Using Laminar Flows of Liquids in Capillaries , 2001 .

[7]  J. Chiu,et al.  A model for studying the effect of shear stress on interactions between vascular endothelial cells and smooth muscle cells. , 2004, Journal of biomechanics.

[8]  Noo Li Jeon,et al.  Patterned cell culture inside microfluidic devices. , 2005, Lab on a chip.

[9]  D. Clark,et al.  Stable sol-gel microstructured and microfluidic networks for protein patterning. , 2001, Biotechnology and bioengineering.

[10]  Heinz Schmid,et al.  Microfluidic Networks for Chemical Patterning of Substrates: Design and Application to Bioassays , 1998 .

[11]  R. Kamm,et al.  Cell migration into scaffolds under co-culture conditions in a microfluidic platform. , 2009, Lab on a chip.

[12]  B. C. Wheeler,et al.  Microstamp patterns of biomolecules for high-resolution neuronal networks , 2006, Medical and Biological Engineering and Computing.

[13]  C. S. Chen,et al.  Geometric control of cell life and death. , 1997, Science.

[14]  R. G. Harrison,et al.  The reaction of embryonic cells to solid structures , 1914 .

[15]  George M. Whitesides,et al.  Features of gold having micrometer to centimeter dimensions can be formed through a combination of stamping with an elastomeric stamp and an alkanethiol ‘‘ink’’ followed by chemical etching , 1993 .

[16]  G M Whitesides,et al.  Patterning cells and their environments using multiple laminar fluid flows in capillary networks. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  G. Owens,et al.  Molecular regulation of vascular smooth muscle cell differentiation in development and disease. , 2004, Physiological reviews.

[18]  Xingyu Jiang,et al.  A method for patterning multiple types of cells by using electrochemical desorption of self-assembled monolayers within microfluidic channels. , 2007, Angewandte Chemie.

[19]  A. Lee,et al.  Engineering microscale cellular niches for three-dimensional multicellular co-cultures. , 2009, Lab on a chip.

[20]  J. Ando,et al.  The effect of flow on the expression of vascular adhesion molecule-1 by cultured mouse endothelial cells. , 1993, Biochemical and biophysical research communications.

[21]  Bruce C Wheeler,et al.  A modified microstamping technique enhances polylysine transfer and neuronal cell patterning. , 2003, Biomaterials.

[22]  T. K. Bhattacharyya,et al.  Microcontact printing of Concanavalin A and its effect on mammalian cell morphology. , 2007, Journal of colloid and interface science.

[23]  D J Beebe,et al.  Microfabricated elastomeric stencils for micropatterning cell cultures. , 2000, Journal of biomedical materials research.

[24]  George M. Whitesides,et al.  Patterning Self-Assembled Monolayers: Applications in Materials Science , 1994 .

[25]  T. Park,et al.  Integration of Cell Culture and Microfabrication Technology , 2003, Biotechnology progress.

[26]  Claire Wyart,et al.  Constrained synaptic connectivity in functional mammalian neuronal networks grown on patterned surfaces , 2002, Journal of Neuroscience Methods.

[27]  R. Timpl,et al.  Neural crest cell migration: requirements for exogenous fibronectin and high cell density , 1983, The Journal of cell biology.

[28]  T. Kodama,et al.  Global analysis of shear stress-responsive genes in vascular endothelial cells. , 2003, Journal of atherosclerosis and thrombosis.

[29]  Matsuhiko Nishizawa,et al.  Controlled cocultures of HeLa cells and human umbilical vein endothelial cells on detachable substrates. , 2009, Lab on a chip.

[30]  R. Nerem.  Vascular fluid mechanics, the arterial wall, and atherosclerosis. , 1992, Journal of biomechanical engineering.

[31]  E. Delamarche,et al.  Patterned delivery of immunoglobulins to surfaces using microfluidic networks. , 1997, Science.

[32]  George M. Whitesides,et al.  Laminar flows: Subcellular positioning of small molecules , 2001, Nature.

[33]  J. Vasiliev,et al.  Behaviour of fibroblast-like cells on grooved surfaces. , 1971, Experimental cell research.

[34]  Witold Dzwinel,et al.  Dynamical clustering of red blood cells in capillary vessels , 2003, Journal of molecular modeling.

[35]  S. Nelander,et al.  DNA microarray study on gene expression profiles in co-cultured endothelial and smooth muscle cells in response to 4- and 24-h shear stress , 2006, Molecular and Cellular Biochemistry.