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Core M focuses on the influence of mechanical stiffness of substrates on the differentiation of stem cells into different lineages. In anchorage-dependent cells, cell adhesion to the extracellular matrix (ECM) is a critical factor that governs cell functions. Hence, the design of biologically responsive materials for regenerative medicine depends on understanding the interactions between cells and the biomaterial/ECM. Among the factors that affect cellular responses, the mechanical stiffness and the presence of adhesive ligands on the biomaterial are known to have considerable effects. The aim of this Core is to characterize the mechanical and adhesive properties of polymer sets and examine/demonstrate their utility in human Mesenchymal Stem Cell differentiation applications. In order to experimentally monitor the lineage of differentiation, this team develops and uses novel lineage-specific, fluorescent-reporter modified stem cells so that the effect of a biomaterial can be measured and quantified. These measurements are then used to further tailor the properties of biomaterials to optimize for a given biomedical application. The team has successfully constructed differentiation reporters in which activation of these promoters drives the expression of a fluorescent marker, either GFP (green fluorescent protein) or mRFP (monomeric red fluorescent protein).
This two-color approach will enable side-by-side evaluation of osteoblastic and adipogenic differentiation in a single experimental setting. These reporter-modified cells are also provided to Core P, which uses multiphoton confocal microscopy to generate cell descriptors during early stages of differentiation. The descriptors are then provided to Core C for computational modeling of the cell-biomaterial interaction. |
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