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Published May 2015 | Supplemental Material
Journal Article Open

A photoreversible protein-patterning approach for guiding stem cell fate in three-dimensional gels

Abstract

Although biochemically patterned hydrogels are capable of recapitulating many critical aspects of the heterogeneous cellular niche, exercising spatial and temporal control of the presentation and removal of biomolecular signalling cues in such systems has proved difficult. Here, we demonstrate a synthetic strategy that exploits two bioorthogonal photochemistries to achieve reversible immobilization of bioactive full-length proteins with good spatial and temporal control within synthetic, cell-laden biomimetic scaffolds. A photodeprotection–oxime-ligation sequence permits user-defined quantities of proteins to be anchored within distinct subvolumes of a three-dimensional matrix, and an ortho-nitrobenzyl ester photoscission reaction facilitates subsequent protein removal. By using this approach to pattern the presentation of the extracellular matrix protein vitronectin, we accomplished reversible differentiation of human mesenchymal stem cells to osteoblasts in a spatially defined manner. Our protein-patterning approach should provide further avenues to probe and direct changes in cell physiology in response to dynamic biochemical signalling.

Additional Information

© 2015 Macmillan Publishers Limited. Received 15 July 2014; Accepted 15 January 2015; Published online 23 February 2015. The authors thank P. Rapp for discussions on FRAP and FEM analysis, as well as for his constructive comments on the manuscript; M. Shahgholi and N. Torian for assistance with HRMS; J. Heath, J. Pfeilsticker and R. Henning for advice on peptide work and for use of their peptide synthesizer and HPLC; D. Koos and the Caltech Biological Imaging Center for use of confocal microscopes; and K. Beres and C. Murry for assistance with all Notch-related studies. This work was supported by the National Science Foundation Grant NSF-DMR 1206121 and a University of Washington Faculty Startup Grant (C.A.D.). Contributions: C.A.D. and D.A.T. designed the experiments. C.A.D. conducted the experiments. C.A.D. and D.A.T. interpreted the data and composed the manuscript. Additional information: Supplementary information is available in the online version of the paper. Competing financial interests: The authors declare no competing financial interests.

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