In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells

Léo Machado (1) , Joana Esteves de Lima (1) , Odile Fabre (2) , Caroline C. Proux (3) , Rachel Legendre (3, 4) , Anikó Szegedi (1) , Hugo Varet (4, 3) , Lars Roed Ingerslev (2) , Romain Barrès (2) , Frédéric Relaix (1) , Philippos Mourikis (1)

1 BNMS - Team 10 - Biologie du système neuromusculaire - Biology of the neuromuscular system [Maisons-Alfort]
2 CBMR - Novo Nordisk Foundation Center for Basic Metabolic Research
3 PF2 - Transcriptome et Epigénome
4 Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB

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.

Keywords: ChIP-seq; RNA-seq; early response genes; methylation; muscle stem cells; quiescence; satellite cells

Domains: Life Sciences [q-bio]; Cellular Biology

Léo Machado, Joana Esteves de Lima, Odile Fabre, Caroline C. Proux, Rachel Legendre, et al.. In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells. Cell Reports, 2017, 21 (7), pp.1982-1993. ⟨10.1016/j.celrep.2017.10.080⟩. ⟨pasteur-02562451⟩