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Systematic Identification of Genes Regulating Muscle Stem Cell Self-Renewal and Differentiation

  • Krishnamoorthy Sreenivasan
  • Thomas Braun
  • Johnny Kim
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1556)

Abstract

The hallmark of stem cells is their capability to either self-renew or to differentiate into a different cell type. Adult skeletal muscle contains a resident muscle stem cell population (MuSCs) known as satellite cells, which enables regeneration of damaged muscle tissue throughout most of adult life. During skeletal muscle regeneration, few MuSCs self-renew to maintain the muscle stem cell pool while others expand rapidly and subsequently undergo myogenic differentiation to form new myofibers. However, like for other stem cell types, the molecular networks that govern self-renewal and/or differentiation of MuSCs remain largely elusive. We recently reported a method to isolate sufficient amounts of purified MuSCs from skeletal muscle which enables us to study their cell autonomous properties. Here, we describe a lentiviral, image-based loss-of function screening pipeline on primary MuSCs that enables systematic identification of genes that regulate muscle stem cell function.

Key words

Muscle stem cells (MuSCs) FAC sorting RNAi screen Lentiviral 

Notes

Acknowledgments

This work was supported by the DFG (Excellence Cluster Cardio-Pulmonary System (ECCPS), Br1416, and SFB TR81), the German-Israeli Fund, and the LOEWE Centre for Cell and Gene Therapy, the German Centre for Cardiovascular Research, and the Universities of Giessen and Marburg Lung Centre (UGMLC).

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Copyright information

© Springer Science+Business Media LLC 2017

Open Access This chapter is distributed under the terms of the Creative Commons Attribution Noncommercial License, which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

Authors and Affiliations

  • Krishnamoorthy Sreenivasan
    • 1
  • Thomas Braun
    • 1
  • Johnny Kim
    • 1
  1. 1.Department of Cardiac Development and RemodellingMax Planck Institute for Heart and Lung ResearchBad NauheimGermany

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