Department of Orthopaedic Surgery, Perelman School of Medicine, The University of Pennsylvania, 112A Stemmler Hall, 3450 Hamilton Walk, Philadelphia, PA 19104-6081, USA.
Development, Aging and Regeneration Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA 92037, USA.
Stem Cell Reports. 2017 Oct 10;9(4):1328-1341. doi: 10.1016/j.stemcr.2017.08.003. Epub 2017 Sep 7.
Muscle stem cells (MuSCs) contribute to muscle regeneration following injury. In many muscle disorders, the repeated cycles of damage and repair lead to stem cell dysfunction. While telomere attrition may contribute to aberrant stem cell functions, methods to accurately measure telomere length in stem cells from skeletal muscles have not been demonstrated. Here, we have optimized and validated such a method, named MuQ-FISH, for analyzing telomere length in MuSCs from either mice or humans. Our analysis showed no differences in telomere length between young and aged MuSCs from uninjured wild-type mice, but MuSCs isolated from young dystrophic mice exhibited significantly shortened telomeres. In corroboration, we demonstrated that telomere attrition is present in human dystrophic MuSCs, which underscores its importance in diseased regenerative failure. The robust technique described herein provides analysis at a single-cell resolution and may be utilized for other cell types, especially rare populations of cells.
肌肉干细胞（MuSCs）在损伤后有助于肌肉再生。在许多肌肉疾病中，损伤和修复的反复循环导致干细胞功能障碍。虽然端粒磨损可能导致异常的干细胞功能，但尚未证明有方法可以准确测量骨骼肌干细胞中的端粒长度。在这里，我们优化并验证了一种名为 MuQ-FISH 的方法，用于分析来自小鼠或人类的 MuSCs 中的端粒长度。我们的分析表明，未受伤的野生型小鼠的年轻和衰老 MuSCs 之间的端粒长度没有差异，但从小龄营养不良型小鼠分离出的 MuSCs 表现出明显缩短的端粒。作为佐证，我们证明了端粒磨损存在于人类营养不良型 MuSCs 中，这凸显了它在疾病性再生失败中的重要性。本文所述的稳健技术提供了单细胞分辨率的分析，可用于其他细胞类型，尤其是稀有细胞群体。
