Regenerative medicine and tissue engineering are being revolutionised by developments in the field of stem cell science. Mesenchymal Stem Cells (MSCs) are emerging as a desirable tool in regenerative medicine and cell therapy due to their wide ranging differentiation potential, large expansion capacity, and their lack of immune rejection following transplantation. Early in vivo studies have demonstrated therapeutic effects of hMSCs; however to clinically exploit the potential of hMSCs, the adherent cell type must be expanded to therapeutically relevant lot sizes (109 to 1012 cells). Hence now there is a need to develop protocols for stable, controlled in vitro expansion, isolation and preservation of a homogenous population of functionally viable cells. Specifically a practical, clinically safe and scalable system which adheres to current GMP guidelines is required to develop reproducible and cost effective therapeutic products. Here we describe the design, manufacture and characterisation of biofunctionalised hydrogel microcarriers containing ECM derived adhesion peptides and a range of compressive moduli for adipose derived stem cell expansion. Microfluidic devices were employed to produce monodisperse spherical particles which were polymerised in situ. In addition, these microcarriers have tunable characteristics which make them a particularly useful tool for the systematic investigation of cellular responses. Microcarriers modified to contain fibronectin and laminin derived peptides supported ADSC attachment and growth in a concentration dependent manner. ADSCs cultured on peptide modified microcarriers were capable of differentiating into osteocytes, chondrocytes and adipocytes, indicating cells cultured on microcarriers maintained multipotency. Substrate compressibility was found to effect ADSC differentiation, corroborating previous literature reports. Bioreactor culture demonstrated successful ADSC expansion with fold increases in cell number far higher than have previously been reported in the literature. High cell seeding densities produced large quantities of viable cells. However, decreasing initial cell seeding density, increased the total fold expansion and reduced cell doubling rates.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:571725 |
| Date | January 2012 |
| Creators | Gibson, Claire |
| Publisher | Cardiff University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://orca.cf.ac.uk/47616/ |
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