This thesis aimed to evaluate the relevancy of different in vitro cell and culture models for osteoblastic-linage cells. Cell lines provide a convenient and accessible alternative to primary human osteoblast cells. However, the direct comparison of these cells demonstrated limited similarity of cell lines to the primary human osteoblasts indicating that their use should be limited to appropriate and specific research questions. To investigate the paracrine regulation of osteogenic development, the immature human osteoblasts and human bone-derived mesenchymal stem cells (MSCs) were co-cultured in monolayer or high density culture. Results from this part of the study suggested the presence of an active signalling pathway between MSCs and osteoblasts. What is more, the effect of cell-cell crosstalk depended on the type of culture system. Co-culture in a 3D micromass system stimulated the osteogenic differentiation of progenitor cells, while in monolayer this was not seen. While the stimulation of MSCs with inflammatory and chemotactic factors successfully regulated the cell gene expression and secretion profile, no effect of the secrotome on the osteogenic differentiation of unstimulated cells in monolayer was demonstrated. Altogether, these results indicated the importance of cell-to-matrix interconnectivity. Therefore, the last part of this thesis focused on the assessment of osteogenic differentiation in 2D and 3D cell culture models, which are physiologically relevant. The progression in osteogenesis depends on the applied 3D culture model. While in both, micromass and type I collagen-hydroxyapatite gel, the differentiation is enhanced compared to monolayer, the regulation of this process is triggered in a different manner in these 3D culture models. Together these findings demonstrate how diverse outcomes can be obtained by the application of different models in in vitro research. Ultimately, the 3D in vitro models provide a better choice for a more in vivo-related osteogenic differentiation and its regulation.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:611035 |
| Date | January 2014 |
| Creators | Czekanska, Ewa Maria |
| Publisher | Cardiff University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://orca.cf.ac.uk/60008/ |
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