Co-culture of SSEA4+-hASCs derived osteoblasts and endothelial cells

• Autor(a) principal: Resende, Margarida Fernandes
• Data de Publicação: 2011
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10773/8710
• topic: Biomedicina; Biologia molecular; Células estaminais; Cultura de células

The crosstalk between Osteoblasts and Endothelial cells is bidirectional and occurs at two levels: indirect cell communication and direct cell communication mediated by proteins at gap junctions structures. The in vitro study of these interactions rely on co-culture systems where at least two cell types are cultured at the same time and submitted to the same microenvironment that is expected to mimic the in vivo settings. Human adipose tissue has been recognized as a potential source of stem cells for regenerative medicine, with multilineage differentiation potential. Recently, it was found that a specific subpopulation of adipose stem cells isolated from the Stromal Vascular Fraction of the Adipose Tissue, positive for the pluripotency associated marker SSEA4 – SSEA4+-hASCs – can give rise to both osteoblastic and microvascular endothelial-like cells, enabling the obtention of these two cell types from the same cell source. This thesis reports the establishment of a co-culture model for the simultaneous culture of two cell types – SSEA4+-hASCs preconditioned osteoblasts (SSEA4+-hASCs OBs) and SSEA4+-hASCs derived endothelial cells. The influence of the cell culture media and the proportion of each cell type influenced the osteogenic and angiogenic outcomes from the system. Our results show that EGM OST culture media allowed to the maintenance of the osteogenic and endothelial differentiation potential of this subpopulation. Concerning the osteogenic and angiogenic potential of the co-cultures, it was shown that both were more consistent on the conditions with higher proportion of ECs in the system. In conclusion, the co-culture condition with the cell ratio OBs:ECs of 1:3 in EGM OST revealed the most efficient osteogenic outcome parameters as well as great stability of endothelial phenotype along the culture period. The established co-culture system has a great potential to overcome the vascularization limitations of the current Bone Tissue Engineering strategies.