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  • Dynamics of Asymmetric and Symmetric Divisions of Muscle Stem Cells In Vivo and on Artificial Niches.

Dynamics of Asymmetric and Symmetric Divisions of Muscle Stem Cells In Vivo and on Artificial Niches.

Cell reports (2020-03-12)
Brendan Evano, Sara Khalilian, Gilles Le Carrou, Geneviève Almouzni, Shahragim Tajbakhsh
ABSTRACT

Stem cells can be maintained through symmetric cell divisions (SCDs) and asymmetric cell divisions (ACDs). How and when these divisions occur in vivo in vertebrates is poorly understood. Here, we developed a clonogenic cell tracing method that demonstrates the asymmetric distribution of transcription factors along with old and new DNA in mouse muscle stem cells during skeletal muscle regeneration. Combining single-cell tracking and artificial niches ex vivo, we show how cells switch from ACDs to SCDs, suggesting that they are not engaged in an obligate mode of cell division. Further, we generated SNAP-tagged histone H3-reporter mice and find that, unlike fly germline stem cells, differential fate outcomes are associated with a symmetric distribution of the H3.1 and H3.3 histone variants in mouse muscle stem cells. This versatile and efficient H3-SNAP labeling system will allow an investigation of mechanisms underlying the maintenance of epigenomic identity and plasticity in a variety of tissues.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonium persulfate, for molecular biology, suitable for electrophoresis, ≥98%
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Triton X-100, laboratory grade
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Acrylamide solution, 40%, suitable for electrophoresis, sterile-filtered
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Fibronectin bovine plasma, solution, sterile-filtered, BioReagent, suitable for cell culture
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N,N,N′,N′-Tetramethylethylenediamine, BioReagent, suitable for electrophoresis, ≥99.0%
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S-Nitroso-N-acetyl-DL-penicillamine, ≥97%, powder
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Tamoxifen, ≥99%
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N,N′-Methylenebisacrylamide solution, suitable for electrophoresis, 2% in H2O
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MCDB 201 Medium, With trace elements, L-glutamine and 30 mM HEPES, powder, suitable for cell culture
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Anti-phospho-Histone H3 (Thr3) Antibody, clone JY325, rabbit monoclonal, clone JY325, from rabbit