Functional immobilization of signaling proteins enables control of stem cell fate

Nat Methods. 2008 Jul;5(7):645-50. doi: 10.1038/nmeth.1222. Epub 2008 Jun 15.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Adhesion
  • Cell Differentiation
  • Cells, Cultured
  • Coated Materials, Biocompatible
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / drug effects
  • Embryonic Stem Cells / metabolism*
  • Leukemia Inhibitory Factor / metabolism*
  • Leukemia Inhibitory Factor / pharmacology
  • Ligands
  • MAP Kinase Signaling System
  • Mice
  • Octamer Transcription Factor-3 / metabolism
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / metabolism*
  • Polymethacrylic Acids
  • Protein Sorting Signals
  • STAT3 Transcription Factor / metabolism
  • Signal Transduction
  • Stem Cell Factor / metabolism

Substances

  • Coated Materials, Biocompatible
  • Leukemia Inhibitory Factor
  • Lif protein, mouse
  • Ligands
  • Octamer Transcription Factor-3
  • Polymethacrylic Acids
  • Pou5f1 protein, mouse
  • Protein Sorting Signals
  • STAT3 Transcription Factor
  • Stat3 protein, mouse
  • Stem Cell Factor
  • poly(n-octyl methacrylate)