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Biosensors 2014, 4(1), 28-44; doi:10.3390/bios4010028

Micropatterning of 3D Microenvironments for Living Biosensor Applications

1 State University of New York (SUNY) College of Nanoscale Science & Engineering, 237 Fuller Road, Albany, NY 12203, USA 2 BioTime, Inc., 1301 Harbor Bay Parkway, Alameda, CA 94502, USA 3 Wisconsin Institutes of Medical Research, University of Wisconsin-Madison, 1111 Highland Ave, Madison, WI 53705, USA These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 24 December 2013 / Revised: 6 February 2014 / Accepted: 17 February 2014 / Published: 27 February 2014
(This article belongs to the Special Issue Sensors and Analytics for Cell Biology and Tissue Engineering)
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Micro-scale printing and patterning of living cells has multiple applications including tissue engineering, cell signaling assays, and the fabrication of cell-based biosensors. In this work, a molecular printing instrument, the Bioforce Nano eNabler, was modified to enable micron-scale “quill-pen” based printing of mammalian cells in a 3D hyaluronan/gelatin based hydrogel. Specifically, photo-initiated “thiol-ene” click chemistry was used to couple the thiol groups of thiolated hyaluronan/thiolated gelatin to the alkene groups of 4-arm polyethylene glycol (PEG)-norbornene molecules. Rapid photopolymerization enabled direct printing and controlled curing of living cells within the hydrogel matrix. The resulting hydrogels were biocompatible with human adipose-derived stem cells, NIH-3T3 cells, and mouse embryonic stem cells. The utility of this printing approach was also explored for cell-based biosensors. Micro-printed cells expressing a redox sensitive variant of the green fluorescent protein (roGFP-R12) showed a measurable fluorescent response to addition of oxidizing and then reducing agents. This work represents a novel approach to micron-scale cell patterning, and its potential for living, cell-based biosensors.
Keywords: microprinting; biosensor; hydrogel; reactive oxygen species; roGFP-R12; ROS; hyaluronan; hyaluronic acid; gelatin; PEG norbornene; Irgacure microprinting; biosensor; hydrogel; reactive oxygen species; roGFP-R12; ROS; hyaluronan; hyaluronic acid; gelatin; PEG norbornene; Irgacure
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Hynes, W.F.; Doty, N.J.; Zarembinski, T.I.; Schwartz, M.P.; Toepke, M.W.; Murphy, W.L.; Atzet, S.K.; Clark, R.; Melendez, J.A.; Cady, N.C. Micropatterning of 3D Microenvironments for Living Biosensor Applications. Biosensors 2014, 4, 28-44.

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