Three-dimensional extracellular matrix-directed cardioprogenitor differentiation: Systematic modulation of a synthetic cell-responsive PEG-hydrogel

• Published in: Biomaterials 2008-06, Vol.29 (18), p.2757-2766
• Main Authors: Kraehenbuehl, Thomas P, Zammaretti, Prisca, Van der Vlies, André J, Schoenmakers, Ronald G, Lutolf, Matthias P, Jaconi, Marisa E, Hubbell, Jeffrey A
• Format: Article
• Language: English
• Subjects: Advanced Basic Science; Animals; Biomimetic hydrogel; Cardiac tissue engineering; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell Line, Tumor; Cell Proliferation - drug effects; Dentistry; Extracellular matrix; Extracellular Matrix - chemistry; Extracellular Matrix - physiology; Flow Cytometry; Hydrogel, Polyethylene Glycol Dimethacrylate - chemical synthesis; Hydrogel, Polyethylene Glycol Dimethacrylate - chemistry; Immunohistochemistry; Muscle, Skeletal - cytology; Muscle, Skeletal - drug effects; Muscle, Skeletal - metabolism; P19 embryonal carcinoma cells; Poly(ethylene glycol); Polyethylene Glycols - chemistry; Reverse Transcriptase Polymerase Chain Reaction; Stem cells; Tissue Engineering - methods
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2008.03.016

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 = 4036 ± 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 (α5 β1, αv β3 ) 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.