Single Stem Cell Imaging and Analysis Reveals Telomere Length Differences in Diseased Human and Mouse Skeletal Muscles.

Single Stem Cell Imaging and Analysis Reveals Telomere Length Differences in Diseased Human and Mouse Skeletal Muscles.
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DOI:
10.1016/j.stemcr.2017.08.003
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发表时间:
2017-10-10
期刊:
影响因子:
5.9
通讯作者:
Mourkioti F
Mourkioti F
中科院分区:
医学1区
文献类型:
--
作者:
Tichy ED;Sidibe DK;Tierney MT;Stec MJ;Sharifi-Sanjani M;Hosalkar H;Mubarak S;Johnson FB;Sacco A;Mourkioti F

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肌肉干细胞(MuSC)有助于损伤后的肌肉再生。在许多肌肉疾病中,损伤和修复的重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,但准确测量骨骼肌干细胞中端粒长度的方法尚未得到证实。在这里,我们已经优化和验证了这种方法,命名为MuQ-FISH,用于分析小鼠或人类MuSC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老年MuSC之间的端粒长度没有差异,但从年轻的营养不良小鼠分离的MuSC表现出显着缩短的端粒。为了证实,我们证明了端粒磨损存在于人类营养不良的MuSC中,这强调了其在疾病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,并且可以用于其他细胞类型,特别是稀有细胞群。MuQ-FISH是小鼠和人类肌肉干细胞的端粒分析测定在少量细胞上的高灵敏度端粒分析使用MuQ-FISH测定检测端粒长度和端粒灶的数量端粒分析现在可以在静止和/或循环干细胞中进行。Alkikioti和同事开发了一种名为MuQ的端粒测定法,FISH结果表明,Duchenne肌营养不良症男孩肌肉干细胞的关键端粒缩短导致了渐进性功能障碍,损害了其再生潜力。
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. MuQ-FISH is a telomere analysis assay of mouse and human muscle stem cells Highly sensitive telomere analysis on small numbers of cells Detection of both telomere length and number of telomere foci with MuQ-FISH assay Telomere analysis is now possible in quiescent and/or cycling stem cells In this article, Mourkioti and colleagues develop a telomere assay named MuQ-FISH and show that the critical telomere shortening of muscle stem cells from boys with Duchenne muscular dystrophy contributes to the progressive dysfunction that compromises their regenerative potential.
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