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In situ cell manipulation through enzymatic hydrogel photopatterning


The physicochemical properties of hydrogels can be manipulated in both space and time through the controlled application of a light beam. However, methods for hydrogel photopatterning either fail to maintain the bioactivity of fragile proteins and are thus limited to short peptides, or have been used in hydrogels that often do not support three-dimensional (3D) cell growth. Here, we show that the 3D invasion of primary human mesenchymal stem cells can be spatiotemporally controlled by micropatterning the hydrogel with desired extracellular matrix (ECM) proteins and growth factors. A peptide substrate of activated transglutaminase factor XIII (FXIIIa)—a key ECM crosslinking enzyme—is rendered photosensitive by masking its active site with a photolabile cage group. Covalent incorporation of the caged FXIIIa substrate into poly(ethylene glycol) hydrogels and subsequent laser-scanning lithography affords highly localized biomolecule tethering. This approach for the 3D manipulation of cells within gels should open up avenues for the study and manipulation of cell signalling.

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Figure 1: Concept of light-controlled enzymatic biomolecule patterning of hydrogels.
Figure 2: Proof-of-principle of light-controlled enzymatic reactions using a soluble model system.
Figure 3: Spatial patterning of hydrogels with fluorescently labelled model ligands and proteins.
Figure 4: In situ manipulation of 3D MSC invasion by light-activated enzymatic patterning.


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We thank A. Negro for help with data analysis, A. Ranga, S. Allazetta, N. Brandenberg, Y. Okawa, K. Krishnamani, P. Abdel-Sayed and N. Balashubrmaniam for valuable discussion, C. Dessibourg, P. Briquez and the Protein Expression Core Facility of EPFL for assistance with recombinant protein production, A. Seitz and T. Laroche for support with confocal microscopy, R. Guiet and O. Burri for assistance with image processing, and S. Banala for sharing his experience with photocaging systems. This work was financially supported in part by a European Young Investigator (EURYI) Award (PE002-117115/1) and an ERC starting grant to M.P.L.

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M.P.L., K.A.M. and L.K. designed research, analysed data and wrote the paper; K.A.M. and L.K. performed research; A.J.v.d.V. contributed to synthesis, purification and analysis of caged molecules; M.M.M., P.S.L., J.A.H. and M.E. contributed new reagents/analytic tools. All authors gave input on the manuscript draft.

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Correspondence to Matthias P. Lutolf.

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Mosiewicz, K., Kolb, L., van der Vlies, A. et al. In situ cell manipulation through enzymatic hydrogel photopatterning. Nature Mater 12, 1072–1078 (2013).

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