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  • Optimisation of procedures for the isolation, culture and characterisation of bovine aortic endothelial cells, human endothelial cells, bovine aortic smooth muscle cells and bovine retinal pericytes

Optimisation of procedures for the isolation, culture and characterisation of bovine aortic endothelial cells, human endothelial cells, bovine aortic smooth muscle cells and bovine retinal pericytes

Procedures have been optimised for the isolation and culture of bovine and human endothelial cells, smooth muscle cells and pericytes. These cells have been plated onto three-dimensional PLGA scaffolds, CaP ceramics and PLGA-CaP composite scaffolds and maintained in static culture, in dynamic cultures and in bioreactors for up to 6 weeks. Using cell biology assays, biochemical assays, SEM analysis and immunohistochemical staining, we have shown that these cells adhere to the scaffolds, proliferate and deposit an extensive extracellular matrix containing type IV collagen, fibronectin and thrombospondin.

However, marked contraction of the PLGA scaffolds was observed when either pericytes alone or co-cultures of pericytes or smooth muscle cells and endothelial cells were plated on the scaffolds, indicating that this scaffold is not suitable for use in bioreactors or in vivo. This contraction was not observed when cells were plated on CaP ceramics or on PLGA-CaP composite scaffolds, demonstrating that the PLGA scaffolds had been strengthened by modification with CaP.

Human endothelial cells did not attach, proliferate or form a contact-inhibited monolayer on these scaffolds to the same extent as bovine cells, suggesting that conditions still need to be optimised for the growth of these cells on these scaffolds.

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University of Manchester
Faculty of Medical & Human Sciences, Michael Smith Building, Oxford Road,
M13 9PT Manchester
United Kingdom
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