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Three-dimensional extracellular matrix-directed cardioprogenitor differentiation: systematic modulation of a synthetic cell-responsive PEG-hydrogel
|Published in||Biomaterials. 2008, vol. 29, no. 18, p. 2757-2766|
|Abstract||We show that synthetic three-dimensional (3D) matrix metalloproteinase (MMP)-sensitive poly(ethylene glycol) (PEG)-based hydrogels can direct differentiation of pluripotent cardioprogenitors, using P19 embryonal carcinoma (EC) cells as a model, along a cardiac lineage in vitro. In order to systematically probe 3D matrix effects on P19 EC differentiation, matrix elasticity, MMP-sensitivity and the concentration of a matrix-bound RGDSP peptide were modulated. Soft matrices (E=322+/-64.2 Pa, stoichiometric ratio: 0.8), mimicking the elasticity of embryonic cardiac tissue, increased the fraction of cells expressing the early cardiac transcription factor Nkx2.5 around 2-fold compared to embryoid bodies (EB) in suspension. In contrast, stiffer matrices (E=4,036+/-419.6 Pa, stoichiometric ratio: 1.2) decreased the number of Nkx2.5-positive cells significantly. Further indicators of cardiac maturation were promoted by ligation of integrins relevant in early cardiac development (alpha(5)beta(1,) alpha(v)beta(3)) by the RGDSP ligand in combination with the MMP-sensitivity of the matrix, with a 6-fold increased amount of myosin heavy chain (MHC)-positive cells as compared to EB in suspension. This precisely controlled 3D culture system thus may serve as a potential alternative to natural matrices for engineering cardiac tissue structures for cell culture and potentially therapeutic applications.|
|Keywords||Animals — Cell Differentiation/drug effects/ physiology — Cell Line, Tumor — Cell Proliferation/drug effects — Extracellular Matrix/chemistry/ physiology — Flow Cytometry — Hydrogel/chemical synthesis/chemistry — Immunohistochemistry — Muscle, Skeletal/cytology/drug effects/metabolism — Polyethylene Glycols/ chemistry — Reverse Transcriptase Polymerase Chain Reaction — Tissue Engineering/ methods|
|Research group||Mécanismes de différentiation cellulaire des cellules cardiaques (536)|