title: An open-chamber flow-focusing device for focal stimulation of micropatterned cells.
abstract: Microfluidic devices can deliver soluble factors to cell and tissue culture microenvironments with precise spatiotemporal control. However, enclosed microfluidic environments often have drawbacks such as the need for continuous culture medium perfusion which limits the duration of experiments, incongruity between microculture and macroculture, difficulty in introducing cells and tissues, and high shear stress on cells. Here, we present an open-chamber microfluidic device that delivers hydrodynamically focused streams of soluble reagents to cells over long time periods (i.e., several hours). We demonstrate the advantage of the open chamber by using conventional cell culture techniques to induce the differentiation of myoblasts into myotubes, a process that occurs in 7-10 days and is difficult to achieve in closed chamber microfluidic devices. By controlling the flow rates and altering the device geometry, we produced sharp focal streams with widths ranging from 36 μm to 187 μm. The focal streams were reproducible (∼12% variation between units) and stable (∼20% increase in stream width over 10 h of operation). Furthermore, we integrated trenches for micropatterning myoblasts and microtraps for confining single primary myofibers into the device. We demonstrate with finite element method (FEM) simulations that shear stresses within the cell trench are well below values known to be deleterious to cells, while local concentrations are maintained at ∼22% of the input concentration. Finally, we demonstrated focused delivery of cytoplasmic and nuclear dyes to micropatterned myoblasts and myofibers. The open-chamber microfluidic flow-focusing concept combined with micropatterning may be generalized to other microfluidic applications that require stringent long-term cell culture conditions.
authors.affiliation: Department of Bioengineering, University of Washington , Seattle, Washington 98195, USA.
authors.forename: Jonathan W
authors.identifier: 0000-0002-7335-7604
authors.initials: JW
authors.lastname: Cheng
authors.affiliation: Department of Bioengineering, University of Washington , Seattle, Washington 98195, USA.
authors.forename: Tim C
authors.initials: TC
authors.lastname: Chang
authors.affiliation: Department of Bioengineering, University of Washington , Seattle, Washington 98195, USA.
authors.forename: Nirveek
authors.initials: N
authors.lastname: Bhattacharjee
authors.affiliation: Department of Bioengineering, University of Washington , Seattle, Washington 98195, USA.
authors.forename: Albert
authors.initials: A
authors.lastname: Folch