Long-term self-renewal of human pluripotent stem cells on peptide-decorated poly(OEGMA-co-HEMA) brushes under fully defined conditions

Acta Biomater. 2013 Nov;9(11):8840-50. doi: 10.1016/j.actbio.2013.07.017. Epub 2013 Jul 24.

Abstract

Realization of the full potential of human induced pluripotent stem cells (hiPSC) in clinical applications requires the development of well-defined culture conditions for their long-term growth and directed differentiation. This paper describes a novel fully defined synthetic peptide-decorated substrate that supports self-renewal of hiPSC in commercially available xeno-free, chemically defined medium. The Au surface was deposited by a poly(OEGMA-co-HEMA) film, using the surface-initiated polymerization method (SIP) with the further step of carboxylation. The hiPSC generated from umbilical cord mesenchymal cells were successfully cultured for 10 passages on the peptide-tethered poly(OEGMA-co-HEMA) brushes for the first time. Cells maintained their characteristic morphology, proliferation and expressed high levels of markers of pluripotency, similar to the cells cultured on Matrigel™. Moreover, the cell adhesion could be tuned by the pattern and peptide concentration on the substrate. This well-defined, xeno-free and safe substrate, which supports long-term proliferation and self-renewal of hiPSC, will not only help to accelerate the translational perspectives of hiPSC, but also provide a platform to elucidate the underlying molecular mechanisms that regulate stem cell proliferation and differentiation via SIP technology.

Keywords: Human pluripotent stem cell; Long-term; Peptide; Self-renewal; Surface-initiated polymerization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocompatible Materials / pharmacology*
  • Cell Adhesion / drug effects
  • Cell Culture Techniques
  • Cell Proliferation / drug effects
  • Cell Shape / drug effects
  • Colony-Forming Units Assay
  • Fluorescence
  • Gene Expression Regulation / drug effects
  • Gold / chemistry
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Immobilized Proteins / pharmacology
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Microscopy, Atomic Force
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Peptides / pharmacology*
  • Polyethylene Glycols / chemistry
  • Polyethylene Glycols / pharmacology*
  • Polymethacrylic Acids / chemistry
  • Polymethacrylic Acids / pharmacology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • SOXB1 Transcription Factors / genetics
  • SOXB1 Transcription Factors / metabolism
  • Surface Properties
  • Time Factors
  • Vitronectin / chemistry
  • Water / chemistry

Substances

  • Biocompatible Materials
  • Homeodomain Proteins
  • Immobilized Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Peptides
  • Polymethacrylic Acids
  • RNA, Messenger
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Vitronectin
  • Water
  • Polyethylene Glycols
  • Gold