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

    Authors

    X. F. Guo; K. Greene; N. Akanda; A. S. T. Smith; M. Stancescu; S. Lambert; H. Vandenburgh;J. J. Hickman

    Comments

    Authors: contact us about adding a copy of your work at STARS@ucf.edu

    Abbreviated Journal Title

    Biomater. Sci.

    Keywords

    RAT SPINAL-CORD; NEUROMUSCULAR-JUNCTION FORMATION; SERUM-FREE MEDIUM; HUMAN-MUSCLE; ELECTRICAL-ACTIVITY; SARCOPLASMIC-RETICULUM; MUSCULAR-DYSTROPHY; TRANSVERSE TUBULES; MESSENGER-RNA; SUBUNIT GENE; Materials Science, Biomaterials

    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.

    Journal Title

    Biomaterials Science

    Volume

    2

    Issue/Number

    1

    Publication Date

    1-1-2014

    Document Type

    Article

    Language

    English

    First Page

    131

    Last Page

    138

    WOS Identifier

    WOS:000330177000014

    ISSN

    2047-4830

    Share

    COinS