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A low resistance microfluidic system for the creation of stable concentration gradients in a defined 3D microenvironment

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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.

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Acknowledgements

This work was supported by National Institute of Health Grants EB003805, AG032977, T32EB006348, R01 AG032977, R37 NS054364, and F31HL095342.

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Authors and Affiliations

  1. Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA

    Ovid C. Amadi

  2. Cardiovascular Division, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA

    Ovid C. Amadi, Matthew L. Steinhauser & Richard T. Lee

  3. Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA

    Yuichi Nishi & Andrew P. McMahon

  4. School of Mechanical Engineering, Korea University, Seoul, South Korea

    Seok Chung

  5. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Roger D. Kamm

  6. Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    Roger D. Kamm

  7. Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA

    Andrew P. McMahon & Richard T. Lee

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  1. Ovid C. Amadi
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  2. Matthew L. Steinhauser
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  3. Yuichi Nishi
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Correspondence to Richard T. Lee.

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Fig. S1

Fig. S1 Rearranged RC circuits used to calculate the time constants for pressure dissipation in the RC device. (a) This circuit represents the path from the source well to the sink well that passes through the reservoir channels and the source and sink reservoir. (b) This circuit represents the path from the source well to sink well that passes through the high resistance gel region (JPEG 318 kb)

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Amadi, O.C., Steinhauser, M.L., Nishi, Y. et al. A low resistance microfluidic system for the creation of stable concentration gradients in a defined 3D microenvironment. Biomed Microdevices 12, 1027–1041 (2010). https://doi.org/10.1007/s10544-010-9457-7

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