title: A low resistance microfluidic system for the creation of stable concentration gradients in a defined 3D microenvironment.
abstract: 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.
authors.affiliation: Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: Ovid C
authors.initials: OC
authors.lastname: Amadi
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Steinhauser
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Nishi
authors.forename: Seok
authors.initials: S
authors.lastname: Chung
authors.forename: Roger D
authors.initials: RD
authors.lastname: Kamm
authors.forename: Andrew P
authors.initials: AP
authors.lastname: McMahon
authors.forename: Richard T
authors.initials: RT
authors.lastname: Lee
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D000077214:Becaplermin; D002417:Cattle; D002454:Cell Differentiation; D002465:Cell Movement; D002633:Chemotaxis; D025802:Electric Capacitance; D017097:Electric Impedance; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D053823:Hedgehog Proteins / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000378:metabolism; D011312:Pressure; D020574:Proto-Oncogene Proteins c-sis; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D000077214:Becaplermin; D053823:Hedgehog Proteins; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis