Micropatterning of 3D Microenvironments for Living Biosensor Applications

Biosensors

William F. Hynes¹  Nate J. Doty³  Thomas I. Zarembinski³  Michael P. Schwartz²  Michael W. Toepke²  William L. Murphy²  Sarah K. Atzet³  Ryan Clark¹  J. Andres Melendez¹ 

[1] State University of New York (SUNY) College of Nanoscale Science & Engineering, 237 Fuller Road, Albany, NY 12203, USA; E-Mails:;Wisconsin Institutes of Medical Research, University of Wisconsin-Madison, 1111 Highland Ave, Madison, WI 53705, USA; E-Mails:;BioTime, Inc., 1301 Harbor Bay Parkway, Alameda, CA 94502, USA; E-Mails:

关键词: microprinting;    biosensor;    hydrogel;    reactive oxygen species;    roGFP-R12;    ROS;    hyaluronan;    hyaluronic acid;    gelatin;    PEG norbornene;    Irgacure;   

DOI  :  10.3390/bios4010028

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.