A low resistance microfluidic system for the creation of stable concentration gradients in a defined 3D microenvironment
- 625 Downloads
The advent of microfluidic technology allows control and interrogation of cell behavior by defining the local microenvironment with an assortment of biochemical and biophysical stimuli. Many approaches have been developed to create gradients of soluble factors, but the complexity of such systems or their inability to create defined and controllable chemical gradients has limited their widespread implementation. Here we describe a new microfluidic device which employs a parallel arrangement of wells and channels to create stable, linear concentration gradients in a gel region between a source and a sink well. Pressure gradients between the source and sink wells are dissipated through low resistance channels in parallel with the gel channel, thus minimizing the convection of solute in this region. We demonstrate the ability of the new device to quantitate chemotactic responses in a variety of cell types, yielding a complete profile of the migratory response and representing the total number of migrating cells and the distance each cell has migrated. Additionally we show the effect of concentration gradients of the morphogen Sonic hedgehog on the specification of differentiating neural progenitors in a 3-dimensional matrix.
KeywordsMicrofluidic Concentration gradient Migration Chemotaxis Morphogen gradient Morphogenesis
This work was supported by National Institute of Health Grants EB003805, AG032977, T32EB006348, R01 AG032977, R37 NS054364, and F31HL095342.
- K.E. Bornfeldt, E.W. Raines, T. Nakano, L.M. Graves, E.G. Krebs, R. Ross, Insulin-like growth factor-I and platelet-derived growth factor-BB induce directed migration of human arterial smooth muscle cells via signaling pathways that are distinct from those of proliferation. J. Clin. Invest. 93(3), 1266–1274 (1994)CrossRefGoogle Scholar
- B.G. Chung, A. Manbachi et al., A gradient-generating microfluidic device for cell biology. J. Vis. Exp. 7, 271 (2007)Google Scholar
- J. Hesselgesser, M. Liang et al., Identification and characterization of the CXCR4 chemokine receptor in human T cell lines: ligand binding, biological activity, and HIV-1 infectivity. J. Immunol. 160(2), 877–883 (1998)Google Scholar
- N. Li Jeon, H. Baskaran et al., Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device. Nat. Biotech. 20(8), 826–830 (2002)Google Scholar
- R.D. Nelson, P.G. Quie et al., Chemotaxis under agarose: a new and simple method for measuring chemotaxis and spontaneous migration of human polymorphonuclear leukocytes and monocytes. J. Immunol. 115(6), 1650–1656 (1975)Google Scholar
- S. Sun, J. Wise et al., Human fibroblast migration in three-dimensional collagen gel in response to noninvasive electrical stimulus. I. Characterization of induced three-dimensional cell movement. Tissue Eng. 10(9–10), 1548–1557 (2004)Google Scholar
- F. Ulloa, E. MartÌ, Wnt won the war: antagonistic role of Wnt over Shh controls dorso-ventral patterning of the vertebrate neural tube. Dev. Dyn. 239(1), 69–76 (2010)Google Scholar
- S.A. Vokes, H. Ji et al., Genomic characterization of Gli-activator targets in sonic hedgehog-mediated neural patterning. Development 134, 1977–1989 (2007)Google Scholar
- S. Wang, J.M. Tarbell, Effect of fluid flow on smooth muscle cells in a 3-dimensional collagen gel model. Arterioscler. Thromb. Vasc. Biol. 20(10), 2220–2225 (2000)Google Scholar
- D. Zicha, G.A. Dunn et al., A new direct-viewing chemotaxis chamber. J. Cell Sci. 99(4), 769–775 (1991)Google Scholar