title: Isolation and Culture of Satellite Cells from Mouse Skeletal Muscle.
abstract: Skeletal muscle tissue is characterized by a population of quiescent mononucleated myoblasts, localized between the basal lamina and sarcolemma of myofibers, known as satellite cells. Satellite cells play a pivotal role in muscle homeostasis and are the major source of myogenic precursors in mammalian muscle regeneration.This chapter describes protocols for isolation and culturing satellite cells isolated from mouse skeletal muscles. The classical procedure, which will be discussed extensively in this chapter, involves the enzymatic dissociation of skeletal muscles, while the alternative method involves isolation of satellite cells from isolated myofibers in which the satellite cells remain in their in situ position underneath the myofiber basal lamina.In particular, we discuss the technical aspect of satellite cell isolation, the methods necessary to enrich the satellite cell fraction and the culture conditions that optimize proliferation and myotube formation of mouse satellite cells.
authors.affiliation: Istituto Pasteur-Fondazione Cenci Bolognetti,DAHFMO-unit of Histology and Medical Embryology, Interuniversity Institute of Myology, Sapienza University of Rome, Via A. Scarpa14, Rome, 00161, Italy. antonio.musaro@uniroma1.it.|Center for Life Nano Science@Sapienza, IstitutoItaliano di Tecnologia, Viale Regina Elena, 291, Rome, 00161, Italy. antonio.musaro@uniroma1.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Musarò
authors.affiliation: Center for Life Nano Science@Sapienza, IstitutoItaliano di Tecnologia, Viale Regina Elena, 291, Rome, 00161, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Carosio
mesh_terms: D000818:Animals; D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D002642:Chick Embryo; D015925:Cryopreservation / Q000379:methods; D018189:Immunomagnetic Separation / Q000379:methods; D016130:Immunophenotyping; D051379:Mice; D008856:Microscopy, Fluorescence; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers
keywords: Cell culture; Muscle differentiation; Muscle primary culture; Myogenic program; Satellite cells isolation