Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation

Yuji Matsuyoshi, Mariko Akahoshi, Mako Nakamura, Ryuichi Tatsumi, Wataru Mizunoya

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

LanguageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages81-93
Number of pages13
DOIs
Publication statusPublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1889
ISSN (Print)1064-3745

Fingerprint

Density Gradient Centrifugation
Cell Separation
Skeletal Muscle
Skeletal Muscle Satellite Cells
Primary Cell Culture
Desmin
Hypertrophy
Regeneration
Stem Cells
Immunohistochemistry
Percoll
Research

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Matsuyoshi, Y., Akahoshi, M., Nakamura, M., Tatsumi, R., & Mizunoya, W. (2019). Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation. In Methods in Molecular Biology (pp. 81-93). (Methods in Molecular Biology; Vol. 1889). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8897-6_6

Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation. / Matsuyoshi, Yuji; Akahoshi, Mariko; Nakamura, Mako; Tatsumi, Ryuichi; Mizunoya, Wataru.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 81-93 (Methods in Molecular Biology; Vol. 1889).

Research output: Chapter in Book/Report/Conference proceedingChapter

Matsuyoshi, Y, Akahoshi, M, Nakamura, M, Tatsumi, R & Mizunoya, W 2019, Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1889, Humana Press Inc., pp. 81-93. https://doi.org/10.1007/978-1-4939-8897-6_6
Matsuyoshi Y, Akahoshi M, Nakamura M, Tatsumi R, Mizunoya W. Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 81-93. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8897-6_6
Matsuyoshi, Yuji ; Akahoshi, Mariko ; Nakamura, Mako ; Tatsumi, Ryuichi ; Mizunoya, Wataru. / Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 81-93 (Methods in Molecular Biology).
@inbook{e80507fa1c384e71a22146db87c90063,
title = "Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation",
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.",
author = "Yuji Matsuyoshi and Mariko Akahoshi and Mako Nakamura and Ryuichi Tatsumi and Wataru Mizunoya",
year = "2019",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-8897-6_6",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "81--93",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Isolation and purification of satellite cells from young rats by percoll density gradient centrifugation

AU - Matsuyoshi, Yuji

AU - Akahoshi, Mariko

AU - Nakamura, Mako

AU - Tatsumi, Ryuichi

AU - Mizunoya, Wataru

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85055612608&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055612608&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-8897-6_6

DO - 10.1007/978-1-4939-8897-6_6

M3 - Chapter

T3 - Methods in Molecular Biology

SP - 81

EP - 93

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -