Andreia Ribeiro
National University of Ireland, Ireland
Title: Effect of extracellular matrix and hypoxia on mesenchymal stromal cell differentiation
Biography
Biography: Andreia Ribeiro
Abstract
Extracellular matrix (ECM) plays an important role in the tumour microenvironment and in biologic processes such as hematopoiesis. ECM contributes to regulation of cell survival, proliferation and cell differentiation. The aims of this project were i) to study the quality, quantity and biological role of ECM produced by a cloned mouse mesenchymal stromal cell line (MS5) on cell differentiation ii) to study the role of hypoxia on cell differentiation. To carry out these studies, we have used two methods of producing ECM in vitro. In both methods ECM is produced in normoxia and hypoxia. In method 1, cells are lysed by osmotic shock with a Tris/EDTA buffer, the standard way of preparing ECM in many studies. In method 2, MS5 were transduced with a caspase 9 vector, allowing induction of apoptosis in the cells following ECM production. Balb/c bone marrow mesenchymal stromal cells (MSC) were then seeded either in uncoated plastic dishes or in dishes covered with ECM and differentiation assays were performed, again either in normoxia or hypoxia. Results show that the two methods produce qualitatively different ECM and that hypoxia plays a role in ECM composition. Moreover, compared with hypoxia, normoxia is a better condition for adipogenic differentiation of fresh MSC. In contracts, osteogenic differentiation is better on ECM in hypoxia. In conclusion, different methods of preparing ECM in vitro lead to different protein composition and different outcomes in cell differentiation. Hypoxia also makes a difference in ECM composition and cell differentiation.