Immobilization of vitronectin-binding heparan sulfates onto surfaces to support human pluripotent stem cells

Lynn Yap, Sadasivam Murali, Gajadhar Bhakta, Drew M. Titmarsh, Allen Kuan Liang Chen, Lyn Chiin Sim, Muriel Bardor, Yu Ming Lim, James C.H. Goh, Steve K.W. Oh, Andre B.H. Choo, Andre J. van Wijnen, David E. Robinson, Jason D. Whittle, William R. Birch, Robert D. Short, Victor Nurcombe, Simon M. Cool

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Functionalizing medical devices with polypeptides to enhance their performance has become important for improved clinical success. The extracellular matrix (ECM) adhesion protein vitronectin (VN) is an effective coating, although the chemistry used to attach VN often reduces its bioactivity. In vivo, VN binds the ECM in a sequence-dependent manner with heparan sulfate (HS) glycosaminoglycans. We reasoned therefore that sequence-based affinity chromatography could be used to isolate a VN-binding HS fraction (HS9) for use as a coating material to capture VN onto implant surfaces. Binding avidity and specificity of HS9 were confirmed by enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR)-based assays. Plasma polymerization of allylamine (AA) to tissue culture-treated polystyrene (TCPS) was then used to capture and present HS9 as determined by radiolabeling and ELISA. HS9-coated TCPS avidly bound VN, and this layered surface supported the robust attachment, expansion, and maintenance of human pluripotent stem cells. Compositional analysis demonstrated that 6-O- and N-sulfation, as well as lengths greater than three disaccharide units (dp6) are critical for VN binding to HS-coated surfaces. Importantly, HS9 coating reduced the threshold concentration of VN required to create an optimally bioactive surface for pluripotent stem cells. We conclude that affinity-purified heparan sugars are able to coat materials to efficiently bind adhesive factors for biomedical applications.

Original languageEnglish (US)
Pages (from-to)1887-1896
Number of pages10
JournalJournal of Biomedical Materials Research - Part B Applied Biomaterials
Volume106
Issue number5
DOIs
StatePublished - Jul 2018

Keywords

  • allylamine
  • biomaterial
  • extracellular matrix
  • glycosaminoglycan
  • implant
  • stem cells
  • surface coating

ASJC Scopus subject areas

  • Biomaterials
  • Biomedical Engineering

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    Yap, L., Murali, S., Bhakta, G., Titmarsh, D. M., Chen, A. K. L., Chiin Sim, L., Bardor, M., Lim, Y. M., Goh, J. C. H., Oh, S. K. W., Choo, A. B. H., van Wijnen, A. J., Robinson, D. E., Whittle, J. D., Birch, W. R., Short, R. D., Nurcombe, V., & Cool, S. M. (2018). Immobilization of vitronectin-binding heparan sulfates onto surfaces to support human pluripotent stem cells. Journal of Biomedical Materials Research - Part B Applied Biomaterials, 106(5), 1887-1896. https://doi.org/10.1002/jbm.b.33999