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

Nat Mater. 2015 May;14(5):523-31. doi: 10.1038/nmat4219. Epub 2015 Feb 23.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biomimetic Materials / chemistry*
  • Cell Differentiation*
  • Humans
  • Hydrogels / chemistry*
  • Immobilized Proteins / chemistry*
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism*
  • Osteoblasts / cytology
  • Osteoblasts / metabolism*
  • Photochemical Processes
  • Tissue Scaffolds / chemistry
  • Vitronectin / chemistry


  • Hydrogels
  • Immobilized Proteins
  • Vitronectin