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
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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.
Hynes WF, Doty NJ, Zarembinski TI, Schwartz MP, Toepke MW, Murphy WL, Atzet SK, Clark R, Melendez JA, Cady NC. Micropatterning of 3D Microenvironments for Living Biosensor Applications. Biosensors. 2014; 4(1):28-44.
Hynes, William F.; Doty, Nate J.; Zarembinski, Thomas I.; Schwartz, Michael P.; Toepke, Michael W.; Murphy, William L.; Atzet, Sarah K.; Clark, Ryan; Melendez, J. A.; Cady, Nathaniel C. 2014. "Micropatterning of 3D Microenvironments for Living Biosensor Applications." Biosensors 4, no. 1: 28-44.