Journal: Stem Cells International

Article Title: VEGF Contributes to Mesenchymal Stem Cell-Mediated Reversion of Nor1-Dependent Hypertrophy in iPS Cell-Derived Cardiomyocytes

doi: 10.1155/2021/8888575

Figure Lengend Snippet: MSC-derived soluble mediators induce hypertrophy regression in iPS-CM. Bone marrow-derived MSCs were stimulated with 30 ng/ml IFN- γ and 3 ng/ml IL-1 β for 24 h. Then, preconditioned MSCs (acMSCS) were indirectly cocultured with hypertrophied iPS-CM. In some experiments, hypertrophied iPS-CM were cultured in medium supplemented with 20% MSC-conditioned medium (CM). (a) Cell area quantification in iPS-CM was performed by immunofluorescent staining. Representative images are displayed. Bar: 100 μ m. n = 6. (b) Nor1 protein expression in iPS-CM. n = 5. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: For experiments, MSCs were stimulated with 30 ng/ml recombinant murine IFN- γ (PeproTech) and 3 ng/ml recombinant murine IL-1 β (PeproTech) in MSC culture medium (Pan-Biotech) supplemented with 2.5 ng/ml human basic fibroblast growth factor FGF (FGF-b, PeproTech), 100 U/ml penicillin, and 10 μ g/ml streptomycin (Sigma-Aldrich) for 24 h. Preconditioned MSCs possess anti-inflammatory and immunomodulatory capacities [ ] and were used for coculture experiments.

Techniques: Derivative Assay, Cell Culture, Staining, Expressing

Journal: Stem Cells International

Article Title: VEGF Contributes to Mesenchymal Stem Cell-Mediated Reversion of Nor1-Dependent Hypertrophy in iPS Cell-Derived Cardiomyocytes

doi: 10.1155/2021/8888575

Figure Lengend Snippet: MSC-derived VEGF is essential for hypertrophy regression in iPS-CM. (a) VEGF was quantified in culture supernatants of MSCs stimulated with 30 ng/ml IFN- γ and 3 ng/ml IL-1 β for 24 h and those cultured for additional 24 h in serum-free medium (48 h). In addition, VEGF levels in fractionated MSC-conditioned medium (concentrates and filtrates) were determined by ELISA. n = 5. (b) PE-treated hypertrophied iPS-CM were cultured in the presence of 20% MSC-conditioned medium (CM) for 24 h. As a control, cells were cultured in medium supplemented with >30 kDa concentrates. For VEGF neutralization, 50 ng/ml or 150 ng/ml neutralizing anti-VEGF antibodies was added to the culture medium. The cell surface area was quantified by F-actin staining. Representative images are displayed. Bar: 100 μ m. n = 5. (c) Nor1 gene and protein expression was quantified in iPS-CM cultured in the presence of MSC-CM supplemented with VEGF-neutralizing antibodies. n = 4. (d) Intracellular VEGF levels were quantified in hypertrophied iPS-CM cultured in the presence of 20% CM for 24 h. MSCs with and without preconditioning served as a control. n = 4. (e) VEGF levels were quantified in culture supernatants of PE-treated iPS-CM cultured in medium supplemented with 20% CM for 24 h. n = 6. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: For experiments, MSCs were stimulated with 30 ng/ml recombinant murine IFN- γ (PeproTech) and 3 ng/ml recombinant murine IL-1 β (PeproTech) in MSC culture medium (Pan-Biotech) supplemented with 2.5 ng/ml human basic fibroblast growth factor FGF (FGF-b, PeproTech), 100 U/ml penicillin, and 10 μ g/ml streptomycin (Sigma-Aldrich) for 24 h. Preconditioned MSCs possess anti-inflammatory and immunomodulatory capacities [ ] and were used for coculture experiments.

Techniques: Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Neutralization, Staining, Expressing