TY - JOUR
T1 - Advanced Methodology for Rapid Isolation of Single Myofibers from Flexor Digitorum Brevis Muscle
AU - Awad, Kamal
AU - Moore, Logan
AU - Huang, Jian
AU - Gomez, Lauren
AU - Brotto, Leticia
AU - Varanasi, Venu
AU - Cardozo, Christopher
AU - Weisleder, Noah
AU - Pan, Zui
AU - Zhou, Jingsong
AU - Bonewald, Lynda
AU - Brotto, Marco
N1 - Publisher Copyright:
© 2023 Mary Ann Liebert Inc.. All rights reserved.
PY - 2023/8/1
Y1 - 2023/8/1
N2 - Isolated individual myofibers are valuable experimental models that can be used in various conditions to understand skeletal muscle physiology and pathophysiology at the tissue and cellular level. This report details a time- and cost-effective method for isolation of single myofibers from the flexor digitorum brevis (FDB) muscle in both young and aged mice. The FDB muscle was chosen for its documented history in single myofiber experiments. By modifying published methods for FDB myofiber isolation, we have optimized the protocol by first separating FDB muscle into individual bundles before the digestion, followed by optimizing the subsequent digestion medium conditions to ensure reproducibility. Morphological and functional assessments demonstrate a high yield of isolated FDB myofibers with sarcolemma integrity achieved in a shorter time frame than previous published procedures. This method could be also adapted to other types of skeletal muscle. Additionally, this highly reproducible method can greatly reduce the number of animals needed to yield adequate numbers of myofibers for experiments. Thus, this advanced method for myofiber isolation has the potential to accelerate research in skeletal muscle physiology and screening potential therapeutics ‘‘ex vivo’’ for muscle diseases and regeneration.
AB - Isolated individual myofibers are valuable experimental models that can be used in various conditions to understand skeletal muscle physiology and pathophysiology at the tissue and cellular level. This report details a time- and cost-effective method for isolation of single myofibers from the flexor digitorum brevis (FDB) muscle in both young and aged mice. The FDB muscle was chosen for its documented history in single myofiber experiments. By modifying published methods for FDB myofiber isolation, we have optimized the protocol by first separating FDB muscle into individual bundles before the digestion, followed by optimizing the subsequent digestion medium conditions to ensure reproducibility. Morphological and functional assessments demonstrate a high yield of isolated FDB myofibers with sarcolemma integrity achieved in a shorter time frame than previous published procedures. This method could be also adapted to other types of skeletal muscle. Additionally, this highly reproducible method can greatly reduce the number of animals needed to yield adequate numbers of myofibers for experiments. Thus, this advanced method for myofiber isolation has the potential to accelerate research in skeletal muscle physiology and screening potential therapeutics ‘‘ex vivo’’ for muscle diseases and regeneration.
KW - FDB
KW - flexor digitorum brevis
KW - isolation
KW - muscle physiology
KW - myofiber
UR - http://www.scopus.com/inward/record.url?scp=85167480222&partnerID=8YFLogxK
U2 - 10.1089/ten.tec.2023.0012
DO - 10.1089/ten.tec.2023.0012
M3 - Article
C2 - 37097213
AN - SCOPUS:85167480222
SN - 1937-3384
VL - 29
SP - 349
EP - 360
JO - Tissue Engineering - Part C: Methods
JF - Tissue Engineering - Part C: Methods
IS - 8
ER -