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

Micropatterning of 3D Microenvironments for Living Biosensor Applications

1,†
,
2,†
,
2
,
3
,
3
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3
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2
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1
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1
 and
1,*
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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Abstract

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 (CC BY 3.0).
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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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