Hybrid Photopatterned Enzymatic Reaction (HyPER) for in Situ Cell Manipulation

• Type of Material: Article
• Authors: Donald R Griffin, University of California, Los Angeles; Jacob Borrajo, University of California, Los Angeles; Allyson Soon, University of California, Los Angeles; Giovanny F. Acosta-Velez, University of California, Los Angeles; Victor Oshita, University of California, Los Angeles; Nicole Darling, University of California, Los Angeles; Julia Mack, University of California, Los Angeles; Thomas Barker, Emory University; M. Luisa Iruela-Arispe, University of California, Los Angeles; Tatiana Segura, University of California, Los Angeles
• Language: English
• Date: 2014-01-24
• Publisher: Wiley: 12 months
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
• Final Published Version (URL): http://dx.doi.org/10.1002/cbic.201300687
• Title of Journal or Parent Work: ChemBioChem
• ISSN: 1439-4227
• Volume: 15
• Issue: 2
• Start Page: 233
• End Page: 242
• Grant/Funding Information: This project was supported by the National Institutes of Health R01NS079691 (TS); and the DermSTP Training Grant T32-AR058921 (DG).
• Keywords: photochemistry; Life Sciences & Biomedicine; STEM-CELLS; HYALURONIC-ACID HYDROGELS; factor XIIIa; FACTOR XIIIA; EXPRESSION; Biochemistry & Molecular Biology; enzyme catalysis; Chemistry, Medicinal; PHOTODEGRADABLE HYDROGELS; nitrobenzene; hydrogels; EXTRACELLULAR-MATRIX; Science & Technology; GROWTH-FACTORS; FIBRIN; PEPTIDES; Pharmacology & Pharmacy; CROSS-LINKING
• Research Categories: Engineering, Chemical; Biology, Cell; Engineering, Biomedical

Abstract

The ability to design artificial extracellular matrices as cell-instructive scaffolds has opened the door to technologies capable of studying the fate of cells in vitro and to guiding tissue repair in vivo. One main component of the design of artificial extracellular matrices is the incorporation of biochemical cues to guide cell phenotype and multicellular organization. The extracellular matrix (ECM) is composed of a heterogeneous mixture of proteins that present a variety of spatially discrete signals to residing cell populations. In contrast, most engineered ECMs do not mimic this heterogeneity. In recent years, photo-deprotection has been used to spatially immobilize signals. However, this approach has been limited mostly to small peptides. Here we combine photo-deprotection with enzymatic reaction to achieve spatially controlled immobilization of active bioactive signals that range from small molecules to large proteins. A peptide substrate for transglutaminase factor XIII (FXIIIa) was caged with a photo-deprotectable group, which was then immobilized to the bulk of a cell-compatible hydrogel. With focused light, the substrate can be deprotected and used to immobilize patterned bioactive signals. This approach offers an innovative strategy to immobilize delicate bioactive signals, such as growth factors, without loss of activity and enables in situ cell manipulation of encapsulated cells. Signal patterns: A peptide substrate has been caged with a photo-deprotectable group, and then immobilized to the bulk of a cell-compatible hydrogel. With focused light, the substrate can be deprotected and used to immobilize patterned bioactive signals without loss of activity and thereby enable in situ cell manipulation of encapsulated cells.