Long term culture of human embryonic stem cells on recombinant vitronectin in ascorbate free media

Andrew B J Prowse, Michael R Doran, Justin J. Cooper-White, Fenny Chong, Trent P Munro, Jane Fitzpatrick, Tung Liang Chung, David N. Haylock, Peter Philip Gray, Ernst J. Wolvetang

Research output: Contribution to journalArticleResearchpeer-review

69 Citations (Scopus)

Abstract

Human embryonic stem cells (hESC) are expected to provide revolutionary therapeutic applications and drug discovery technologies. In order for this to be achieved a reproducible, defined animal component free culture system is required for the scale-up production of undifferentiated hESC. In this work we have investigated the applicability of a recombinantly produced domain of human vitronectin as an extracellular matrix alternative to the common standards Geltrex™ or Matrigel™. In addition we have validated an ascorbate free media capable of supporting CD30low populations of hESC through a multi-factorial analysis of bFGF and Activin A. The recombinant vitronectin domain combined with the ascorbate free media were capable of supporting 3 cell lines, MEL1, MEL2 and hES3 for 10 or more passages while maintaining hESC pluripotency markers and differentiation capacity. The culture method outlined here provides a platform for future investigation into growth factor and extracellular matrix effects on hESC maintenance prior to bioreactor scale-up.

Original languageEnglish
Pages (from-to)8281-8288
Number of pages8
JournalBiomaterials
Volume31
Issue number32
DOIs
Publication statusPublished - Nov 2010
Externally publishedYes

Keywords

  • Bioreactor
  • Cell adhesion
  • Extracellular matrix
  • Growth factor
  • Recombinant protein
  • Stem cell

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