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

:卫星细胞位于体内肌肉中,是支持骨骼肌生长和再生的Pax7肌肉干细胞。不幸的是,其治疗用途所需的体外扩增与降低的再生潜力有关。在本研究中,我们调查了体外获得的人肌原性储备细胞(MRC)是否代表肌肉修复的可靠细胞来源。为此目的,新鲜分离并扩增了原代人成肌细胞。分化2天后,有62%±2.9%的细胞核位于肌管中,有38%±2.9%的细胞位于单核非融合性MRC中。 80%的新鲜分离出的人类MRC表现出与人类静止卫星细胞(CD56 / Pax7 / MyoD- / Ki67-细胞)相似的表型。在免疫缺陷小鼠中,细胞移植后第14天和第21天,活人细胞明显增多,并且与注射人成肌细胞的动物相比,注射MRC的动物的肌肉中Pax7 /人层粘连A / C细胞的百分比高2倍,尽管两组中的血影蛋白和层状A / C人纤维百分比均相似。综上所述,这些数据提供了证据,证明体外产生的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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