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Myogenesis pp 81-93 | Cite as

Isolation and Purification of Satellite Cells from Young Rats by Percoll Density Gradient Centrifugation

  • Yuji Matsuyoshi
  • Mariko Akahoshi
  • Mako Nakamura
  • Ryuichi Tatsumi
  • Wataru MizunoyaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1889)

Abstract

Satellite cells (SCs) are myogenic stem cells that play an important role in skeletal muscle regeneration and hypertrophy. Primary cultures of SCs are useful to analyze cell functions; however, it is difficult to obtain highly pure SCs from young rats with the conventional procedures. The purpose of this study is to establish a purification method for SC isolation from young rats and quantitatively evaluate the purification procedure employing Percoll, a common research tool to purify cells. We elucidated the purity of SCs collected by Percoll density gradient centrifugation using real-time RT-qPCR and immunocytochemistry for desmin. Percoll treatment increased the purity of SCs isolated from young rats to nearly 90%, which was comparable to that achieved with the conventional method using middle-aged rats.

Key words

Skeletal muscle Cell isolation Cell purification Satellite cells Percoll density gradient centrifugation 

Notes

Acknowledgments

This work was funded by the Japan Society for the Promotion of Science (JSPS) KAKENHI #26712023 to WM. Many thanks to Ms. Akiko Sato and Mr. Shuichi Kitaura (Kyushu University, Japan) for animal care and for help with handling chemical reagents.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yuji Matsuyoshi
    • 1
  • Mariko Akahoshi
    • 1
  • Mako Nakamura
    • 1
  • Ryuichi Tatsumi
    • 1
  • Wataru Mizunoya
    • 1
    Email author
  1. 1.Department of Bioresource Sciences, Faculty of AgricultureKyushu UniversityFukuokaJapan

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