Satellite cells, localized within muscles in vivo, are Pax7+ muscle stem cells supporting skeletal muscle growth and regeneration. Unfortunately, their amplification in vitro, required for their therapeutic use, is associated with reduced regenerative potential. In the present study, we investigated if human myogenic reserve cells (MRC) obtained in vitro, represented a reliable cell source for muscle repair. For this purpose, primary human myoblasts were freshly isolated and expanded. After 2 days of differentiation, 62 ± 2.9% of the nuclei were localized in myotubes and 38 ± 2.9% in the mononucleated non-fusing MRC. Eighty percent of freshly isolated human MRC expressed a phenotype similar to human quiescent satellite cells (CD56+/Pax7+/MyoD-/Ki67- cells). Fourteen days and 21 days after cell transplantation in immunodeficient mice, live human cells were significantly more numerous and the percentage of Pax7+/human lamin A/C+ cells was 2 fold higher in muscles of animals injected with MRC compared to those injected with human myoblasts, despite that percentage of spectrin+ and lamin A/C+ human fibers in both groups MRC were similar. Taken together, these data provide evidence that MRC generated in vitro represent a promising source of cells for improving regeneration of injured skeletal muscles.

译文

体内定位在肌肉内的卫星细胞是Pax7 + 肌肉干细胞,支持骨骼肌生长和再生。不幸的是,其治疗用途所需的体外扩增与再生潜力降低有关。在本研究中,我们研究了体外获得的人肌源性储备细胞 (MRC) 是否代表了用于肌肉修复的可靠细胞来源。为此,新鲜分离并扩增了原代人类成肌细胞。分化2天后,62个nuclei ±   2.9% 的细胞核位于肌管中,38个   ±   2.9% 位于单核非融合MRC中。80% 的新鲜分离的人MRC表达的表型类似于人静止卫星细胞 (CD56/Pax7/MyoD-/Ki67-细胞)。免疫缺陷小鼠细胞移植后14天和21天,活人细胞数量明显增加,注射MRC的动物肌肉中Pax7 +/人lamin A/C + 细胞的百分比比注射人成肌细胞高2倍,尽管两组MRC中spectrin和lamin A/C人类纤维的百分比相似。总之,这些数据提供了证据,表明体外产生的MRC代表了改善受损骨骼肌再生的有希望的细胞来源。

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