He basal lamina (Figure 4B). Similarly, satellite cells grafted in BaCl

He basal lamina (Figure 4B). Similarly, satellite cells grafted in BaCl2injured muscles formed few donor-derived fibres (464) and the presence of donor-derived nuclei inside and outside the fibres was rare (161 and 261 respectively) (Figure 4B, C-II, D-II). BaCl2?treated muscles injected with single fibres rather than those injected with satellite cells were significantly heavier than either BaCl2 reated muscles injected with DMEM, or muscles irradiated and grafted with satellite cells (Figure 4E). The significant increase in CSA in BaCl2 reated muscles injected with single fibres mirrored this difference (Figure 4F). Since the total number of fibres in BaCl2 pre-injured single fibre-grafted muscles was not significantly increased (Figure 3E and Figure 4G), we conclude that the grafted donor fibre plays a pivotal role in promoting the hypertrophic effect in host muscles.DiscussionEvidence that a single grafted donor myofibre can dramatically change host skeletal muscle by contributing robustly to skeletal muscle regeneration came from experiments employing the same in vivo system as we used here ?fibres from donor geneticallymodified wild type mice grafted into pre-irradiated muscles of dystrophin-deficient mdx nude mice [6]. Further studies showed that modulation of the host muscle environment is an important requirement for successful donor satellite cell engraftment: not only does the host niche need to be preserved, but also endogenous satellite cells have to be impaired [45]. Such modulation, achieved by irradiating host muscles, permits aged host muscle to be regenerated by donor satellite cells as well as young host muscle [7,47]. Myotoxins, such as BaCl2, notexin and cardiotoxin, have been widely used to cause muscle I-BRD9 injury [48,49]. These destroy myofibres, but myofibre basal lamina, satellite cells, nerves and blood vessels are preserved [48]. In response to the muscle injury, endogenous satellite cells activate, proliferate, migrate and either get JI 101 repair injured fibres, or regenerate new fibres [50,51]; thus the contribution of transplanted donor cells in competition with efficient host-mediated muscle regeneration is negligible [45]. Among the myotoxins we tested, BaCl2 was the only one, when injected 3 days before cell grafting, that promoted significantly more donor-derived muscle formation than in the non-treated host muscles, even though donor muscle formation was 10 times less than in the irradiated grafted muscles [45]. We 1662274 were therefore interested to see the effect of BaCl2 on grafted single fibres, bearing their complement of satellite cells. We clearly show that, in our model system, donor muscle formation derived from isolated donor myofibres grafted into in BaCl2-injured host mdx nude muscles is rare and insignificant. However, although they do not give rise to either muscle fibres, or other cell types, within BaCl2-treated host muscles, a donor single fibre stimulated host muscle hypertrophy. The number of fibres has not increased, but the diameter of the fibres has, leading to a significant increase in muscle weight. The effect of the grafted isolated fibre on the host muscle is therefore hypertrophy, not hyperplasia, as it is an increase in fibre size rather than number. Intriguingly, this donor fibre-mediated hypertrophic effect occurred without pre-injury of the host muscle with BaCl2, indicating that non-treated mdx nude muscles, which would beThe Hypertrophic Effect is Mediated by the Donor Fibre Rather than Don.He basal lamina (Figure 4B). Similarly, satellite cells grafted in BaCl2injured muscles formed few donor-derived fibres (464) and the presence of donor-derived nuclei inside and outside the fibres was rare (161 and 261 respectively) (Figure 4B, C-II, D-II). BaCl2?treated muscles injected with single fibres rather than those injected with satellite cells were significantly heavier than either BaCl2 reated muscles injected with DMEM, or muscles irradiated and grafted with satellite cells (Figure 4E). The significant increase in CSA in BaCl2 reated muscles injected with single fibres mirrored this difference (Figure 4F). Since the total number of fibres in BaCl2 pre-injured single fibre-grafted muscles was not significantly increased (Figure 3E and Figure 4G), we conclude that the grafted donor fibre plays a pivotal role in promoting the hypertrophic effect in host muscles.DiscussionEvidence that a single grafted donor myofibre can dramatically change host skeletal muscle by contributing robustly to skeletal muscle regeneration came from experiments employing the same in vivo system as we used here ?fibres from donor geneticallymodified wild type mice grafted into pre-irradiated muscles of dystrophin-deficient mdx nude mice [6]. Further studies showed that modulation of the host muscle environment is an important requirement for successful donor satellite cell engraftment: not only does the host niche need to be preserved, but also endogenous satellite cells have to be impaired [45]. Such modulation, achieved by irradiating host muscles, permits aged host muscle to be regenerated by donor satellite cells as well as young host muscle [7,47]. Myotoxins, such as BaCl2, notexin and cardiotoxin, have been widely used to cause muscle injury [48,49]. These destroy myofibres, but myofibre basal lamina, satellite cells, nerves and blood vessels are preserved [48]. In response to the muscle injury, endogenous satellite cells activate, proliferate, migrate and either repair injured fibres, or regenerate new fibres [50,51]; thus the contribution of transplanted donor cells in competition with efficient host-mediated muscle regeneration is negligible [45]. Among the myotoxins we tested, BaCl2 was the only one, when injected 3 days before cell grafting, that promoted significantly more donor-derived muscle formation than in the non-treated host muscles, even though donor muscle formation was 10 times less than in the irradiated grafted muscles [45]. We 1662274 were therefore interested to see the effect of BaCl2 on grafted single fibres, bearing their complement of satellite cells. We clearly show that, in our model system, donor muscle formation derived from isolated donor myofibres grafted into in BaCl2-injured host mdx nude muscles is rare and insignificant. However, although they do not give rise to either muscle fibres, or other cell types, within BaCl2-treated host muscles, a donor single fibre stimulated host muscle hypertrophy. The number of fibres has not increased, but the diameter of the fibres has, leading to a significant increase in muscle weight. The effect of the grafted isolated fibre on the host muscle is therefore hypertrophy, not hyperplasia, as it is an increase in fibre size rather than number. Intriguingly, this donor fibre-mediated hypertrophic effect occurred without pre-injury of the host muscle with BaCl2, indicating that non-treated mdx nude muscles, which would beThe Hypertrophic Effect is Mediated by the Donor Fibre Rather than Don.