TY - JOUR
T1 - Three-dimensional extracellular matrix-directed cardioprogenitor differentiation
T2 - Systematic modulation of a synthetic cell-responsive PEG-hydrogel
AU - Kraehenbuehl, Thomas P.
AU - Zammaretti, Prisca
AU - Van der Vlies, André J.
AU - Schoenmakers, Ronald G.
AU - Lutolf, Matthias P.
AU - Jaconi, Marisa E.
AU - Hubbell, Jeffrey A.
N1 - Funding Information:
We thank Dr. Mayumi Mochizuki and Conlin P. O'Neil for help with peptide synthesis, Dr. Mathieu Hauwel for support with RT-PCR, Miriella Pasquier for assistance in tissue embedding and processing, Olivier Brun for supporting with confocal imaging and Sergei Startchik for helping with confocal image processing. We also thank Prof. Ilona Skerjanc, University of Western Ontario, Canada, and Dr. Marcel Van der Heyden, University Medical Center Utrecht, The Netherlands, for helpful discussions. This work was supported by grants of the Novartis Foundation for Medicine and Biology and of the European Union's 6th framework program Expertissues.
PY - 2008/6
Y1 - 2008/6
N2 - 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 (α5β1, αvβ3) 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.
AB - 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 (α5β1, αvβ3) 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.
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U2 - 10.1016/j.biomaterials.2008.03.016
DO - 10.1016/j.biomaterials.2008.03.016
M3 - Article
C2 - 18396331
AN - SCOPUS:42049085710
SN - 0142-9612
VL - 29
SP - 2757
EP - 2766
JO - Biomaterials
JF - Biomaterials
IS - 18
ER -