Isolation and Culture of Individual Myofibers and Their Adjacent Muscle Stem Cells from Aged and Adult Skeletal Muscle

Methods Mol Biol. 2019:2045:25-36. doi: 10.1007/7651_2019_209.

Abstract

The isolation and culture of single floating myofibers with their adjacent muscle stem cells allow the analysis and comparison of muscle stem cells from aged and young mice. This method has the advantage that muscle stem cells are cultured on the myofiber, thereby culturing them in conditions as close to their endogenous niche as possible. Here we describe the isolation, culture, transfection with siRNA, and subsequent immunostaining for muscle stem cells on their adjacent myofibers from aged and young mice.

Keywords: Aging; Collagenase; Differentiation; Muscle stem cell; Myofiber; Satellite cell; Self-renewal; Transfection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells / cytology*
  • Adult Stem Cells / metabolism
  • Aging
  • Animals
  • Antibodies
  • Cell Culture Techniques / methods*
  • Cell Differentiation
  • Collagenases
  • Immunophenotyping
  • Mice
  • Muscle Fibers, Skeletal / cytology*
  • Muscle Fibers, Skeletal / metabolism
  • Muscle, Skeletal / cytology*
  • Muscle, Skeletal / metabolism
  • MyoD Protein / immunology
  • MyoD Protein / metabolism
  • PAX7 Transcription Factor / immunology
  • PAX7 Transcription Factor / metabolism
  • RNA, Small Interfering
  • Satellite Cells, Skeletal Muscle / cytology
  • Satellite Cells, Skeletal Muscle / metabolism
  • Transfection
  • Workflow

Substances

  • Antibodies
  • MyoD Protein
  • MyoD1 myogenic differentiation protein
  • PAX7 Transcription Factor
  • Pax7 protein, mouse
  • RNA, Small Interfering
  • Collagenases