In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells
In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells
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DOI：
10.1016/j.celrep.2017.10.080
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发表时间：
2017-11-14
期刊：
影响因子：
8.8
通讯作者：
Mourikis, Philippos
中科院分区：
文献类型：
--
作者：
Machado, Leo;de Lima, Joana Esteves;Mourikis, Philippos
已经开发了最先进的技术来分离和分析来自各种组织的细胞，旨在捕获它们的体内状态。然而，大多数细胞分离方案涉及漫长的机械和酶解离步骤，然后进行流式细胞术，使细胞暴露于压力并破坏其生理生态位。专注于成人骨骼肌干细胞，我们已经开发了一个协议，规避隔离程序的影响，并捕获细胞在其天然的静止状态。我们发现，目前的隔离协议诱导主要的转录变化伴随着特定的组蛋白修饰，而对DNA甲基化的影响可以忽略不计。除了提出一个协议，以避免孤立引起的文物，我们的研究揭示了以前未被发现的潜在的生物学兴趣的静止和早期激活基因。
State of the art techniques have been developed to isolate and analyze cells from various tissues, aiming to capture their in vivo state. However, the majority of cell isolation protocols involve lengthy mechanical and enzymatic dissociation steps followed by flow cytometry, exposing cells to stress and disrupting their physiological niche. Focusing on adult skeletal muscle stem cells, we have developed a protocol that circumvents the impact of isolation procedures and captures cells in their native quiescent state. We show that current isolation protocols induce major transcriptional changes accompanied by specific histone modifications while having negligible effects on DNA methylation. In addition to proposing a protocol to avoid isolation-induced artifacts, our study reveals previously undetected quiescence and early activation genes of potential biological interest.