Issue 1, 2014

In vitro differentiation of functional human skeletal myotubes in a defined system

Abstract

In vitro human skeletal muscle systems are valuable tools for the study of human muscular development, disease and treatment. However, published in vitro human muscle systems have so far only demonstrated limited differentiation capacities. Advanced differentiation features such as cross-striations and contractility have only been observed in co-cultures with motoneurons. Furthermore, it is commonly regarded that cultured human myotubes do not spontaneously contract, and any contraction has been considered to originate from innervation. This study developed a serum-free culture system in which human skeletal myotubes demonstrated advanced differentiation. Characterization by immunocytochemistry, electrophysiology and analysis of contractile function revealed these major features: (A) Well defined sarcomeric development, as demonstrated by the presence of cross-striations. (B) Finely developed excitation–contraction coupling apparatus characterized by the close apposition of dihydropyridine receptors on T-tubules and ryanodine receptors on sarcoplasmic reticulum membranes. (C) Spontaneous and electrically controlled contractility. This report not only demonstrates an improved level of differentiation of cultured human skeletal myotubes, but also provides the first published evidence that such myotubes are capable of spontaneous contraction. Use of this functional in vitro human skeletal muscle system would advance studies concerning human skeletal muscle development and physiology, as well as muscle-related disease and therapy.

Graphical abstract: In vitro differentiation of functional human skeletal myotubes in a defined system

Supplementary files

Article information

Article type
Paper
Submitted
02 Jul 2013
Accepted
04 Sep 2013
First published
25 Sep 2013

Biomater. Sci., 2014,2, 131-138

In vitro differentiation of functional human skeletal myotubes in a defined system

X. Guo, K. Greene, N. Akanda, A. S. T. Smith, M. Stancescu, S. Lambert, H. Vandenburgh and J. J. Hickman, Biomater. Sci., 2014, 2, 131 DOI: 10.1039/C3BM60166H

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