Culturing of Cardiac Fibroblasts in Engineered Heart Matrix Reduces Myofibroblast Differentiation but Maintains Their Response to Cyclic Stretch and Transforming Growth Factor β1

• Published in: Bioengineering (Basel) Vol. 9; no. 10; p. 551
• Main Authors: Ploeg, Meike C, Munts, Chantal, Seddiqi, Tayeba, ten Brink, Tim J. L, Breemhaar, Jonathan, Moroni, Lorenzo, Prinzen, Frits. W, van Nieuwenhoven, Frans. A
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.10.2022; MDPI
• Subjects: Actin; Bioengineering; Biomarkers; Brain natriuretic peptide; cardiac fibroblast; Cell culture; Collagen; Collagen (type I); Connective tissue; Connective tissue growth factor; Connective tissues; Differentiation; Extracellular matrix; Fibroblasts; Gene expression; Growth factors; Health aspects; Heart; Hydrogels; Mechanical properties; Modulus of elasticity; Monolayers; Muscles; Peptides; Phenotypes; Smooth muscle; Stiffness; Stimulation; stretch; Substrates; Tensile strength; three dimensional; Three dimensional models; Transforming growth factor-b1; Netherlands; United States > US; Germany
• ISSN: 2306-5354; 2306-5354
• DOI: 10.3390/bioengineering9100551

Isolation and culturing of cardiac fibroblasts (CF) induces rapid differentiation toward a myofibroblast phenotype, which is partly mediated by the high substrate stiffness of the culture plates. In the present study, a 3D model of Engineered Heart Matrix (EHM) of physiological stiffness (Youngs modulus ~15 kPa) was developed using primary adult rat CF and a natural hydrogel collagen type 1 matrix. CF were equally distributed, viable and quiescent for at least 13 days in EHM and the baseline gene expression of myofibroblast-markers alfa-smooth muscle actin (Acta2), and connective tissue growth factor (Ctgf) was significantly lower, compared to CF cultured in 2D monolayers. CF baseline gene expression of transforming growth factor-beta1 (Tgfβ1) and brain natriuretic peptide (Nppb) was higher in EHM-fibers compared to the monolayers. EHM stimulation by 10% cyclic stretch (1 Hz) increased the gene expression of Nppb (3.0-fold), Ctgf (2.1-fold) and Tgfβ1 (2.3-fold) after 24 h. Stimulation of EHM with TGFβ1 (1 ng/mL, 24 h) induced Tgfβ1 (1.6-fold) and Ctgf (1.6-fold). In conclusion, culturing CF in EHM of physiological stiffness reduced myofibroblast marker gene expression, while the CF response to stretch or TGFβ1 was maintained, indicating that our novel EHM structure provides a good physiological model to study CF function and myofibroblast differentiation.