Abstract
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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Acknowledgements
We thank T. Lenk and J. Drichel for technical assistance. P.W.Z. is a Canada Research Chair in Stem Cell Bioengineering. R.E.D. was supported by a Natural Sciences and Engineering Research Council of Canada (NSERC) award. K.O. was supported by an Ontario Graduate Scholarship. This work was accomplished with support from NSERC (Discovery and Steacie). F.P.S., M.B. and C.W. were supported by the Deutsche Forschungsgemeinschaft, “Collaborative Research Centre: cells into tissues—stem cell and progenitor commitment and interactions during tissue formation” (SFB 655). C.W. was supported by the Federal Ministry of Science, Education and Technology of Germany (grant 0N4022 Center of Excellence in Biomaterials, Dresden).
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R.E.D., K.A., K.S., K.O., S.G. and F.P.S. designed and performed experiments and assisted with writing the manuscript. M.B., T.P., P.W.Z., A.N. and C.W. designed the experiments and wrote the manuscript.
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Alberti, K., Davey, R., Onishi, K. et al. Functional immobilization of signaling proteins enables control of stem cell fate. Nat Methods 5, 645–650 (2008). https://doi.org/10.1038/nmeth.1222
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DOI: https://doi.org/10.1038/nmeth.1222
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