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Methods for proteomics-based analysis of the human muscle secretome using an in vitro exercise model.
In: Proteomic Profiling. Berlin [u.a.]: Springer, 2021. 433-442 (Methods Mol. Biol. ; 2261)
Over the last decade, the skeletal muscle as a secretory organ has gained importance. A growing number of peptides is described which are produced and released by the muscle fibers and work in an autocrine, paracrine, and endocrine fashion. The contraction-induced secretion of these myokines is considered to contribute to the health promoting effects of exercise. To gain further insights into the molecular processes that occur during contraction, an in vitro exercise model, electric pulse stimulation (EPS), was established. Recent publications show that this model is suitable to electrostimulate human skeletal muscle cells and thus mimic muscle contraction in vitro. Here, we provide a detailed protocol for the proteomics-based analysis of the human muscle secretome, starting with the cultivation of human myotubes and ending with sample preparation for targeted and untargeted proteome analysis of the cell culture supernatant. This workflow should allow for deeper insights into the complex nature of the muscle secretome and the identification of new myokines which might help to understand the cross talk of the working muscle with different organs and the beneficial effects of exercise.
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Publication type
Article: Edited volume or book chapter
Keywords
Electric Pulse Stimulation ; Human Myotubes ; Lc-ms ; Macs ; Multiplex Immunoassay ; Myokine ; Secretome
ISSN (print) / ISBN
1064-3745
e-ISSN
1940-6029
Book Volume Title
Proteomic Profiling
Journal
Methods in Molecular Biology
Quellenangaben
Volume: 2261,
Pages: 433-442
Publisher
Springer
Publishing Place
Berlin [u.a.]
Non-patent literature
Publications
Reviewing status
Peer reviewed