a-e) Histological sections of aortic valve tissue from male patients diagnosed with heart valve disorder. Immunostaining for fibrotic markers, a) αSMA and b) COL1A1 (green, nucleus in blue), show high variability between tissues and heterogeneity in the fibroblast and myofibroblast population, indicated by αSMA staining. Calcific markers, c) RUNX2 (red, nucleus in blue), d) Von Kossa (calcium phosphate in black) and e) Alizarin Red (calcium in red) demonstrate high variability in tissue mineralization. Scale bars=100μm. Quantification of f) αSMA g) COL1A1 and h) RUNX2 intensity analysis for tissue sections, significant differences are observed between tissues for αSMA and RUNX2. i) Quantification of area percentage of Von Kossa positive stain (black) indicates variability of calcification between tissues. Statistical analysis was performed on fields of view for each biological replicate (n≥10 images per biological replicate), ***=p<0.001 and **=p<0.01 based on one-way ANOVA.