Mesenchymal Stem Cells of Epicardial Adipose Tissue Show Differences in Immunophenotype and Osteogenic Potential in Patients with Coronary and Non-Coronary Heart Disease

Figures

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  Figure 1. Assessment of growth parameters of EAT-MSC cultures from CAD and AS patients. Note: (a) Proliferation curves on days 3, 7, and 10; (b) Specific growth rate of a crop (c) Population doubling time on days 3, 7, and 10; (d) Cell viability on day 3.

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  Figure 2. Micrographs of EAT MSCs stained with β-galactosidase at passage 3, (β-gal)-positive cells are stained blue: (a) EAT MSCs from patients with CAD; (b) EAT MSCs from patients with AS; (c) comparative assessment of the proportion of old cells in the cultures.

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  Figure 3. Detection of RUNX2 in EAT-MSC cultures from CAD and AS patients during osteogenic differentiation. Note: (a) Relative expression level of RUNX2 on days 3, 15, and 21 of osteogenic differentiation; (b) RUNX2 protein levels in culture supernatants; (c) Immunofluorescence staining of RUNX2 protein in EAT-MSCs (intranuclear localization).

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  Figure 4. Detection of osteopontin in EAT-MSC cultures from CAD and AS patients during osteogenic differentiation. Note: (a) Relative expression level of the gene encoding osteopontin (SPP1) on days 3, 15, and 21 of osteogenic differentiation; (b) Osteopontin protein levels in culture supernatants; (c) Immunofluorescence staining of osteopontin protein in EAT-MSCs (intracellular localization).

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  Figure 5. Detection of alkaline phosphatase (ALP) in EAT-MSC cultures from CAD and AS patients during osteogenic differentiation. Note: (a) Relative expression level of the gene encoding ALP (ALPL) on days 3, 15, and 21 of osteogenic differentiation; (b) ALP protein levels in culture supernatants; (c) Immunofluorescence staining of ALP protein in EAT-MSCs (intracellular localization).

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  Figure 6. Detection of osteocalcin in EAT-MSC cultures from CAD and AS patients during osteogenic differentiation. Note: (a) Relative expression level of the gene encoding osteocalcin (BGLAP) on days 3, 15, and 21 of osteogenic differentiation; (b) Osteocalcin protein levels in culture supernatants; (c) Immunofluorescence staining of osteocalcin protein in EAT-MSCs (extracellular localization).

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  Figure 7. Micro- and macrophotographs of Alizarin Red staining of osteoblasts derived from EAT-MSCs of CAD and AS patients, and quantitative measurement of staining intensity. Note: (a) Osteoblasts from EAT-MSCs of CAD patients; (b) Osteoblasts from EAT-MSCs of AS patients; (c) Photometric measurement of calcium-bound Alizarin Red concentration.