Assessing Muscle Stem Cell Clonal Complexity During Aging

Methods Mol Biol. 2019:2045:1-11. doi: 10.1007/7651_2018_139.

Abstract

Changes in muscle stem cell (MuSC) function during aging have been assessed using various in vivo and ex vivo systems. However, changes in clonal complexity within the aged MuSC pool are relatively understudied. Although the dissection of stem cell heterogeneity has greatly benefited from several technological advancements, including single cell sequencing, these methods preclude longitudinal measures of individual stem cell behavior. Instead, multicolor labeling systems enable lineage tracing with single cell resolution. Here, we describe a method of inducibly labeling MuSCs with the Brainbow-2.1 multicolor lineage tracing reporter in vivo to track individual MuSC fate and assess clonal complexity in the overall MuSC pool throughout the mouse lifespan.

Keywords: Aging; Brainbow; Clonal complexity; Multicolor lineage tracing; Satellite cell; Skeletal muscle.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aging / physiology*
  • Animals
  • Cell Lineage
  • Cluster Analysis
  • Cryoultramicrotomy
  • Genes, Reporter / drug effects
  • Genes, Reporter / genetics
  • Luminescent Proteins / analysis*
  • Luminescent Proteins / chemistry*
  • Mice
  • Microscopy, Confocal
  • Muscle, Skeletal / cytology*
  • Myoblasts / cytology*
  • Software
  • Staining and Labeling / methods*
  • Stem Cells / cytology*
  • Workflow

Substances

  • Luminescent Proteins