Nature Methods
- 5, 645 - 650 (2008)
Published online: 15 June 2008; | doi:10.1038/nmeth.1222
Functional immobilization of signaling proteins enables control of stem cell fateKristin Alberti1, 7, Ryan E Davey2, 7, Kento Onishi2, Sophia George3, Katrin Salchert1, F Philipp Seib1, 4, Martin Bornhäuser4, 5, Tilo Pompe1, Andras Nagy3, Carsten Werner1, 2, 5 & Peter W Zandstra2, 61
Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Str. 6, D-01069 Dresden, Germany. 2
Institute of Biomaterials and Biomedical Engineering, 164 College Street, University of Toronto, Toronto, M5S 3G9, Canada. 3
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, M5G 1X5, Canada. 4
University Hospital Carl Gustav Carus, Dresden University of Technology, Fetscherstr. 74, D-01307, Dresden, Germany. 5
Center for Regenerative Therapies Dresden, Tatzberg 47/49, D-01307 Dresden, Germany. 6
Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, M5S 3E5, Canada. 7
These authors contributed equally to this work.
Correspondence should be addressed to Peter W Zandstra peter.zandstra@utoronto.ca or Carsten Werner werner@ipfdd.de The mode of ligand presentation has a fundamental role in organizing cell fate throughout development. We report a rapid and simple approach for immobilizing signaling ligands to maleic anhydride copolymer thin-film coatings, enabling stable signaling ligand presentation at interfaces at defined concentrations. We demonstrate the utility of this platform technology using leukemia inhibitory factor (LIF) and stem cell factor (SCF). Immobilized LIF supported mouse embryonic stem cell (mESC) pluripotency for at least 2 weeks in the absence of added diffusible LIF. Immobilized LIF activated signal transducer and activator of transcription 3 (STAT3) and mitogen-activated protein kinase (MAPK) signaling in a dose-dependent manner. The introduced method allows for the robust investigation of cell fate responses from interface-immobilized ligands.
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