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Human epicardium-derived cells fuse with high efficiency with skeletal myotubes and differentiate toward the skeletal muscle phenotype: a comparison study with stromal and endothelial cells.

Gentile A, Toietta G, Pazzano V, Tsiopoulos VD, Giglio AF, Crea F, Pompilio G, Capogrossi MC, Di Rocco G - Mol. Biol. Cell (2011)

Bottom Line: Results are compared to those obtained with mesenchymal stromal cells (MSCs) and with endothelial cells, another mesodermal derivative.We additionally show that vascular cell adhesion molecule 1 (VCAM1) expression levels of nonmuscle cells are modulated by soluble factors secreted by skeletal myoblasts and that VCAM1 function is required for fusion to occur.Finally, treatment with interleukin (IL)-4 or IL-13, two cytokines released by differentiating myotubes, increases VCAM1 expression and enhances the rate of fusion of EPDCs and MSCs, but not that of endothelial cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio di Patologia Vascolare, Istituto Dermopatico dell'Immacolata-IRCCS, 00167 Rome, Italy.

ABSTRACT
Recent studies have underscored a role for the epicardium as a source of multipotent cells. Here, we investigate the myogenic potential of adult human epicardium-derived cells (EPDCs) and analyze their ability to undergo skeletal myogenesis when cultured with differentiating primary myoblasts. Results are compared to those obtained with mesenchymal stromal cells (MSCs) and with endothelial cells, another mesodermal derivative. We demonstrate that EPDCs spontaneously fuse with pre-existing myotubes with an efficiency that is significantly higher than that of other cells. Although at a low frequency, endothelial cells may also contribute to myotube formation. In all cases analyzed, after entering the myotube, nonmuscle nuclei are reprogrammed to express muscle-specific genes. The fusion competence of nonmyogenic cells in vitro parallels their ability to reconstitute dystrophin expression in mdx mice. We additionally show that vascular cell adhesion molecule 1 (VCAM1) expression levels of nonmuscle cells are modulated by soluble factors secreted by skeletal myoblasts and that VCAM1 function is required for fusion to occur. Finally, treatment with interleukin (IL)-4 or IL-13, two cytokines released by differentiating myotubes, increases VCAM1 expression and enhances the rate of fusion of EPDCs and MSCs, but not that of endothelial cells.

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Transplantation of EPDCs and MSCs but not endothelial cells rescues dystrophin expression in mdx mice. TA transverse sections from transplanted mdx mice. Staining for dystrophin (red) was performed 7 d after injection. Nuclei are visualized by Hoechst (blue). Several clusters of dystrophin-positive fibers can be detected in mice that received injections with EPDCs and MSCs, whereas the number of dystrophin-positive fibers detected with endothelial cells (ENDO) does not goes beyond the number of revertant fibers present in uninjected controls. Magnification: 40×, Bar: 25 μm.
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Figure 4: Transplantation of EPDCs and MSCs but not endothelial cells rescues dystrophin expression in mdx mice. TA transverse sections from transplanted mdx mice. Staining for dystrophin (red) was performed 7 d after injection. Nuclei are visualized by Hoechst (blue). Several clusters of dystrophin-positive fibers can be detected in mice that received injections with EPDCs and MSCs, whereas the number of dystrophin-positive fibers detected with endothelial cells (ENDO) does not goes beyond the number of revertant fibers present in uninjected controls. Magnification: 40×, Bar: 25 μm.

Mentions: To further assess the myogenic differentiation potential of EPDCs, we wanted to investigate whether EPDCs can be recruited to form skeletal myofibers in vivo. EPDCs cultured for not more than two passages were injected into the tibialis anterior (TA) muscle of pharmacologically immunosuppressed mdx mice. As in the previous experiment, AT-MSCs and endothelial cells at P2 were used for comparison. Ten days after injection, less than 0.1% dystrophin-positive fibers, corresponding to spontaneously revertant fibers, were observed in sections from age-matched phosphate-buffered saline (PBS)-injected mice. Conversely, dystrophin was detected in up to 10% (10.3 ± 2.3% in three different experiments) of the myofibers analyzed, corresponding to 1% of the entire area, on sections from EPDC-transplanted muscles, as revealed by immunofluorescence with an antibody against the C-terminal portion of mouse dystrophin (Figure 4). Dystrophin-positive fibers were organized in clusters, suggesting clonal proliferation of donor cells. Analogous results, although with a reduced number of dystrophin-positive fibers (5.7 ± 1.5%, approximately 0.5% of the entire area), were obtained with AT-MSCs. Conversely, with endothelial cells the number of dystrophin-positive fibers was never higher than that of spontaneous revertants, corresponding to 0.1 ± 0.05% fibers (<0.01% of the area) in the 2-mo-old mdx mice used for our experiments.


Human epicardium-derived cells fuse with high efficiency with skeletal myotubes and differentiate toward the skeletal muscle phenotype: a comparison study with stromal and endothelial cells.

Gentile A, Toietta G, Pazzano V, Tsiopoulos VD, Giglio AF, Crea F, Pompilio G, Capogrossi MC, Di Rocco G - Mol. Biol. Cell (2011)

Transplantation of EPDCs and MSCs but not endothelial cells rescues dystrophin expression in mdx mice. TA transverse sections from transplanted mdx mice. Staining for dystrophin (red) was performed 7 d after injection. Nuclei are visualized by Hoechst (blue). Several clusters of dystrophin-positive fibers can be detected in mice that received injections with EPDCs and MSCs, whereas the number of dystrophin-positive fibers detected with endothelial cells (ENDO) does not goes beyond the number of revertant fibers present in uninjected controls. Magnification: 40×, Bar: 25 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 4: Transplantation of EPDCs and MSCs but not endothelial cells rescues dystrophin expression in mdx mice. TA transverse sections from transplanted mdx mice. Staining for dystrophin (red) was performed 7 d after injection. Nuclei are visualized by Hoechst (blue). Several clusters of dystrophin-positive fibers can be detected in mice that received injections with EPDCs and MSCs, whereas the number of dystrophin-positive fibers detected with endothelial cells (ENDO) does not goes beyond the number of revertant fibers present in uninjected controls. Magnification: 40×, Bar: 25 μm.
Mentions: To further assess the myogenic differentiation potential of EPDCs, we wanted to investigate whether EPDCs can be recruited to form skeletal myofibers in vivo. EPDCs cultured for not more than two passages were injected into the tibialis anterior (TA) muscle of pharmacologically immunosuppressed mdx mice. As in the previous experiment, AT-MSCs and endothelial cells at P2 were used for comparison. Ten days after injection, less than 0.1% dystrophin-positive fibers, corresponding to spontaneously revertant fibers, were observed in sections from age-matched phosphate-buffered saline (PBS)-injected mice. Conversely, dystrophin was detected in up to 10% (10.3 ± 2.3% in three different experiments) of the myofibers analyzed, corresponding to 1% of the entire area, on sections from EPDC-transplanted muscles, as revealed by immunofluorescence with an antibody against the C-terminal portion of mouse dystrophin (Figure 4). Dystrophin-positive fibers were organized in clusters, suggesting clonal proliferation of donor cells. Analogous results, although with a reduced number of dystrophin-positive fibers (5.7 ± 1.5%, approximately 0.5% of the entire area), were obtained with AT-MSCs. Conversely, with endothelial cells the number of dystrophin-positive fibers was never higher than that of spontaneous revertants, corresponding to 0.1 ± 0.05% fibers (<0.01% of the area) in the 2-mo-old mdx mice used for our experiments.

Bottom Line: Results are compared to those obtained with mesenchymal stromal cells (MSCs) and with endothelial cells, another mesodermal derivative.We additionally show that vascular cell adhesion molecule 1 (VCAM1) expression levels of nonmuscle cells are modulated by soluble factors secreted by skeletal myoblasts and that VCAM1 function is required for fusion to occur.Finally, treatment with interleukin (IL)-4 or IL-13, two cytokines released by differentiating myotubes, increases VCAM1 expression and enhances the rate of fusion of EPDCs and MSCs, but not that of endothelial cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio di Patologia Vascolare, Istituto Dermopatico dell'Immacolata-IRCCS, 00167 Rome, Italy.

ABSTRACT
Recent studies have underscored a role for the epicardium as a source of multipotent cells. Here, we investigate the myogenic potential of adult human epicardium-derived cells (EPDCs) and analyze their ability to undergo skeletal myogenesis when cultured with differentiating primary myoblasts. Results are compared to those obtained with mesenchymal stromal cells (MSCs) and with endothelial cells, another mesodermal derivative. We demonstrate that EPDCs spontaneously fuse with pre-existing myotubes with an efficiency that is significantly higher than that of other cells. Although at a low frequency, endothelial cells may also contribute to myotube formation. In all cases analyzed, after entering the myotube, nonmuscle nuclei are reprogrammed to express muscle-specific genes. The fusion competence of nonmyogenic cells in vitro parallels their ability to reconstitute dystrophin expression in mdx mice. We additionally show that vascular cell adhesion molecule 1 (VCAM1) expression levels of nonmuscle cells are modulated by soluble factors secreted by skeletal myoblasts and that VCAM1 function is required for fusion to occur. Finally, treatment with interleukin (IL)-4 or IL-13, two cytokines released by differentiating myotubes, increases VCAM1 expression and enhances the rate of fusion of EPDCs and MSCs, but not that of endothelial cells.

Show MeSH
Related in: MedlinePlus