Protocol for rat single muscle fiber isolation and culture.
To attain a superior in vitro model of mature muscle fibers, we modified the established protocol for isolating single muscle fibers from rat skeletal muscle. Muscle fiber cultures with high viability were obtained using flexor digitorum brevis muscle and lasted for at least 7 days. We compared the expression levels of adult myosin heavy chain (MyHC) isoforms in these single muscle fibers with myotubes formed from myoblasts; isolated fibers contained markedly more abundant adult MyHC isoforms than myotubes. This muscle fiber model, therefore, will be useful for studying the various functions and cellular processes of mature muscles in vitro.
Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan.
Yusuke
Y
Komiya
Department of Biological Sciences, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada.
Judy E
JE
Anderson
Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan.
Mariko
M
Akahoshi
Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan.
Mako
M
Nakamura
Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan.
Ryuichi
R
Tatsumi
Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan.
Yoshihide
Y
Ikeuchi
Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan. Electronic address: wataru_m@agr.kyushu-u.ac.jp.
Wataru
W
Mizunoya
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D008297:Male; D018485:Muscle Fibers, Skeletal / Q000166:cytology*; D018482:Muscle, Skeletal / Q000166:cytology*; D032446:Myoblasts / Q000166:cytology; D018995:Myosin Heavy Chains / Q000032:analysis; D020033:Protein Isoforms / Q000032:analysis; D051381:Rats; D011916:Rats, Inbred F344
D020033:Protein Isoforms; D018995:Myosin Heavy Chains
Muscle fiber; Primary culture; Rat; Skeletal muscle