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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.

Authors :
Machado L
Esteves de Lima J
Fabre O
Proux C
Legendre R
Szegedi A
Varet H
Ingerslev LR
Barrès R
Relaix F
Mourikis P
Source :
Cell reports [Cell Rep] 2017 Nov 14; Vol. 21 (7), pp. 1982-1993.
Publication Year :
2017

Abstract

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.<br /> (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
2211-1247
Volume :
21
Issue :
7
Database :
MEDLINE
Journal :
Cell reports
Publication Type :
Academic Journal
Accession number :
29141227
Full Text :
https://doi.org/10.1016/j.celrep.2017.10.080