Journal: Stem Cells International

Article Title: Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells

doi: 10.1155/2015/498328

Figure Lengend Snippet: Characterization of BM-MSCs: detailed FACS characterization revealed that MSCs at P3 to P5 stained negatively for CD14 and CD45 but expressed surface markers that are indicative of MSC lineage, including CD44, CD73, and CD90 (a). Isolated MSCs also exhibited fibroblastoid-like morphology (b). Naïve BM-MSCs demonstrated the capacity to differentiate in osteogenic, chondrogenic, and adipogenic lineages. Alizarin red showed staining of calcium deposits in MSCs differentiated into the osteogenic lineage. Alcian/Safranin O blue showed staining of peptidoglycans characteristic of differentiation into the chondrogenic lineage. Oil Red O showed staining of the lipids and triglycerides, indicating differentiation into the adipogenic lineage. Each image shown is representative of independent experiments performed with BM-MSC cultures derived from separate microswine ( n = 3). FACS analysis performed for EC markers on naïve MSCs (c), MSCs treated with VEGF-A (d), and HUVECs (e). Each grid shown is representative of independent experiments performed with cultures derived from bone marrow of separate microswine ( n = 3–6).

Article Snippet: Yucatan microswine (30–40 lbs) was purchased from Sinclair Bioresources (Windham, Maine).

Techniques: Staining, Isolation, Derivative Assay

Journal: Stem Cells International

Article Title: Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells

doi: 10.1155/2015/498328

Figure Lengend Snippet: Angioplasty increases the local production of atherogenic cytokines: histological evaluation was performed on LCX arteries taken from hypercholesterolemic microswine at 4 months after angioplasty and age-matched control hypercholesterolemic microswine. The immunostaining was assessed for Ang II ((a)-(b)), TNF α ((c)-(d)), and IL-6 ((e)-(f)). Each image shown is representative of independent experiments performed from separate microswine ( n = 3).

Article Snippet: Yucatan microswine (30–40 lbs) was purchased from Sinclair Bioresources (Windham, Maine).

Techniques: Control, Immunostaining

Journal: Stem Cells International

Article Title: Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells

doi: 10.1155/2015/498328

Figure Lengend Snippet: IL-6 negatively regulates EC differentiation: the effect of IL-6 on VEGF-A-stimulated differentiation of MSCs into ECs was examined. IL-6R and VEGFR-2 mRNA expression was analyzed by RT-PCR and normalized to GAPDH ( n = 3) (a). Sox18 protein levels were measured by Western blot analysis and normalized to GAPDH ( n = 3) (b). Expression of EC markers was determined by FACS analysis, and a representative grid is shown ( n = 3–6) ((c)–(g)). Endothelial tube formation was examined using an angiogenesis assay ( n = 3) ((d)–(h)). Experiments were performed with samples taken from independent BM-MSC cultures from separate microswine. HUVECs were excluded from statistical analyses. Data are shown as mean ± SD. ∗ p < 0.05 versus naïve MSCs and # p < 0.05 MSCs treated with VEGF-A versus VEGF-A plus IL-6 cotreatment.

Article Snippet: Yucatan microswine (30–40 lbs) was purchased from Sinclair Bioresources (Windham, Maine).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Angiogenesis Assay

Journal: Stem Cells International

Article Title: Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells

doi: 10.1155/2015/498328

Figure Lengend Snippet: TNF α negatively regulates EC differentiation: the effect of TNF α on VEGF-A-stimulated differentiation of MSCs into ECs was examined. TNF α R and VEGFR-2 mRNA expression was analyzed by RT-PCR ( n = 3) (a). Sox18 protein levels were measured by Western blot analysis ( n = 3) (b). Expression of EC markers was determined by FACS analysis, and a representative grid is shown ( n = 3-4) ((c)–(g)). Endothelial tube formation was examined using an angiogenesis assay ( n = 3) ((d)–(h)). Experiments were performed with samples taken from independent BM-MSC cultures from separate microswine. HUVECs were excluded from statistical analyses. Data are shown as mean ± SD. ∗ p < 0.05 versus naïve MSCs and # p < 0.05 MSCs treated with VEGF-A versus VEGF-A plus TNF α cotreatment.

Article Snippet: Yucatan microswine (30–40 lbs) was purchased from Sinclair Bioresources (Windham, Maine).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Angiogenesis Assay

Journal: Stem Cells International

Article Title: Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells

doi: 10.1155/2015/498328

Figure Lengend Snippet: Ang II positively regulates VEGF-A-mediated EC differentiation: the effect of Ang II on VEGF-A-stimulated differentiation of MSCs into ECs was examined. AT2R and VEGFR-2 mRNA expression was analyzed by RT-PCR ( n = 3) (a). Sox18 protein levels were measured by Western blot analysis ( n = 3) (b). Expression of EC markers was determined by FACS analysis, and a representative grid is shown ( n = 5-6) ((c)–(g)). Endothelial tube formation was examined using an angiogenesis assay ( n = 3) ((d)–(h)). Experiments were performed with samples taken from independent BM-MSC cultures from separate microswine. HUVECs were excluded from statistical analyses. Data are shown as mean ± SD. ∗ p < 0.05 versus naïve MSCs and # p < 0.05 MSCs treated with VEGF-A versus VEGF-A plus Ang II cotreatment.

Article Snippet: Yucatan microswine (30–40 lbs) was purchased from Sinclair Bioresources (Windham, Maine).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Angiogenesis Assay

Journal: Stem Cells International

Article Title: Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells

doi: 10.1155/2015/498328

Figure Lengend Snippet: Ang II counteracts IL-6 or TNF α inhibition of EC differentiation: the combined effects of Ang II, IL-6, and TNF α on VEGF-A-stimulated differentiation of MSCs into ECs were examined. TNF α R, IL-6R, AT2R, and VEGFR-2 mRNA expression was analyzed by RT-PCR ( n = 3) (a). Sox18 protein levels were measured by Western blot analysis ( n = 3) (b). Expression of EC markers was determined by FACS analysis, and a representative grid is shown ( n = 3-4) ((c)–(h)). Endothelial tube formation was examined using an angiogenesis assay ( n = 3) ((d)–(i)). Experiments were performed with samples taken from independent BM-MSC cultures from separate microswine. HUVECs were excluded from statistical analyses. Data are shown as mean ± SD. ∗ p < 0.05 versus naïve MSCs, # p < 0.05 MSCs treated with VEGF-A versus VEGF-A plus Ang II cotreatment, and α p < 0.05 versus MSCs treated with VEGF-A and Ang II versus VEGF-A and Ang II plus IL-6 and/or TNF α .

Article Snippet: Yucatan microswine (30–40 lbs) was purchased from Sinclair Bioresources (Windham, Maine).

Techniques: Inhibition, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Angiogenesis Assay