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[Jove]A Versatile Method of Patterning Proteins and Cells


Abstract:

Substrate and cell patterning techniques are widely used in cell biology to study cell-to-cell and cell-to-substrate interactions. Conventional patterning techniques work well only with simple shapes, small areas and selected bio-materials. This article describes a method to distribute cell suspensions as well as substrate solutions into complex, long, closed (dead-end) polydimethylsiloxane (PDMS) microchannels using negative pressure. This method enables researchers to pattern multiple substrates including fibronectin, collagen, antibodies (Sal-1), poly-D-lysine (PDL), and laminin. Patterning of substrates allows one to indirectly pattern a variety of cells. We have tested C2C12 myoblasts, the PC12 neuronal cell line, embryonic rat cortical neurons, and amphibian retinal neurons. In addition, we demonstrate that this technique can directly pattern fibroblasts in microfluidic channels via brief application of a low vacuum on cell suspensions. The low vacuum does not significantly decrease cell viability as shown by cell viability assays. Modifications are discussed for application of the method to different cell and substrate types. This technique allows researchers to pattern cells and proteins in specific patterns without the need for exotic materials or equipment and can be done in any laboratory with a vacuum.

Shrirao, A. B., Kung, F. H., Yip, D., Firestein, B. L., Cho, C. H., Townes-Anderson, E. A Versatile Method of Patterning Proteins and Cells. J. Vis. Exp. (120), e55513, doi:10.3791/55513 (2017).

Anil B. Shrirao*1, Frank H. Kung*2, Derek Yip3, Bonnie L. Firestein2, Cheul H, Cho3, Ellen Townes-Anderson4

1Department of Biomedical Engineering, Rutgers University,

2Department of Cell Biology and Neuroscience, Rutgers University,

3Department of Biomedical Engineering, New Jersey Institute of Technology,

4Department of Pharmacology, Physiology, and Neuroscience, Rutgers New Jersey Medical School

Shrirao, A. B., Kung, F. H., Yip, D., Firestein, B. L., Cho, C. H., Townes-Anderson, E. A Versatile Method of Patterning Proteins and Cells. J. Vis. Exp. (120), e55513, doi:10.3791/55513 (2017).

Link: https://www.jove.com/video/55513/a-versatile-method-of-patterning-proteins-and-cells

#03142017 #patterning #cellpatterning #substrate #Biologicalapplication

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