Long term mesenchymal stem cell culture on a defined synthetic substrate with enzyme free passaging

Cairnan R.E. Duffy, Rong Zhang, Siew-Eng How, Annamaria Lilienkampf, Paul A. De Sousa, Mark Bradley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Mesenchymal stems cells (MSCs) are currently the focus of numerous therapeutic approaches in tissue engineering/repair because of their wide multi-lineage potential and their ability to modulate the immune system response following transplantation. Culturing these cells, while maintaining their multipotency in vitro, currently relies on biological substrates such as gelatin, collagen and fibronectin. In addition, harvesting cells from these substrates requires enzymatic or chemical treatment, a process that will remove a multitude of cellular surface proteins, clearly an undesirable process if cells are to be used therapeutically. Herein, we applied a high-throughput 'hydrogel microarray' screening approach to identify thermo-modulatable substrates which can support hES-MP and ADMSC growth, permit gentle reagent free passaging, whilst maintaining multi-lineage potential. In summary, the hydrogel substrate identified, poly(AEtMA-Cl-co-DEAA) cross-linked with MBA, permitted MSCs to be maintained over 10 passages (each time via thermo-modulation), with the cells retaining expression of MSC associated markers and lineage potency. This chemically defined system allowed the passaging and maintenance of cellular phenotype of this clinically important cell type, in the absence of harsh passaging and the need for biological substrates.

Original languageEnglish
Pages (from-to)5998-6005
Number of pages8
JournalBiomaterials
Volume35
Issue number23
Early online date26 Apr 2014
DOIs
Publication statusPublished - 31 Jul 2014

Keywords

  • Mesenchymal stem cell
  • Hydrogel
  • Polymer microarray
  • Cell culture
  • MARROW STROMAL CELLS
  • MATRIX-MEDIATED RETENTION
  • HUMAN UMBILICAL-CORD
  • EX-VIVO EXPANSION
  • MICROARRAYS
  • HYDROGEL
  • POLYMERS
  • BLOOD

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