Myofiber Isolation: A Technique to Obtain Single Myofibers from Harvested Mouse Extensor Digitorum Longus (EDL) Muscle

Abstract

Source: Hüettner, S.S. et al. Single Myofiber Culture Assay for the Assessment of Adult Muscle Stem Cell Functionality Ex Vivo. J. Vis. Exp. (2021)

This video demonstrates the isolation of single myofibres from harvested murine Extensor Digitorum Longus (EDL) muscle. The isolated myofibres can be cultured to study myofiber-associated muscle stem cell activity.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. EDL muscle isolation and collagenase digestion

1. Spray all equipment with 70% ethanol to avoid contamination.
2. Sacrifice the mouse in accordance with the national regulations for animal experimentation.
3. Spray the hindlimbs of the mouse with 70% ethanol. Use hardened fine curved scissors (24 mm cutting edge) and fine forceps (Dumont 7, curved or straight) to remove the skin and to expose the underlying muscles. Avoid any contact of the underlying musc....

Results

Figure 1. Mouse EDL dissection and single myofiber isolation. (A) The skin of the mouse hindlimb is removed and the muscle surrounding fascia is pulled away to expose the TA (tibialis anterior) muscle. (

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Collagenase type 1	Sigma	C0130
DMEM (Dulbecco’s modified Eagle’s medium with 4.5 g/l glucose and sodium pyruvate)	GibCo	41966029	Cell culture medium
Fetal bovine serum	Gibco	10270-106	Fetal bovine serum