Embryonic motoneuron-skeletal muscle co-culture in a defined system

    Authors

    M. Das; J. W. Rumsey; C. A. Gregory; N. Bhargava; J. F. Kang; P. Molnar; L. Riedel; X. Guo;J. J. Hickman

    Comments

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    Abbreviated Journal Title

    Neuroscience

    Keywords

    defined system; neuromuscular junction; self-assembled monolayer; motoneuron; serum-free medium; skeletal muscle; SERUM-FREE MEDIUM; RAT SPINAL-CORD; NEUROMUSCULAR-JUNCTION; IN-VITRO; ACETYLCHOLINE-RECEPTOR; ORGANOSILANE SURFACE; HIPPOCAMPAL-NEURONS; CULTURE; GROWTH; CELLS; Neurosciences

    Abstract

    This paper describes a significant biotechnological advancement by creating a minimalist serum-free defined system to co-culture rat mammalian nerve and muscle cells in order to form functional neuromuscular junctions. To date, all the known in vitro nerve and muscle co-culture models use serum containing media; and while functional neuromuscular junctions (NMJ) are described, they failed to detail or quantify the minimum factors needed to recreate the NMJ in vitro. In this work, we demonstrate the development of a defined motoneuron and muscle co-culture system resulting in the formation of NMJs including: 1) a new culture technique, 2) a novel serum-free medium formulation and 3) a synthetic self-assembled monolayer (SAM) substrate N-1 [3-(trimethoxysilyi) propyl] diethylenetriamine (DETA). We characterized the culture by morphology, immunocytochemistry, electrophysiology and videography. This model system provides a better understanding of the minimal growth factor and substrate interactions necessary for NMJ formation and provides a basic system that can be utilized for nerve-muscle tissue engineering, regenerative medicine and development of limb prosthetics. (C) 2007 IBRO. Published by Elsevier Ltd. All rights reserved.

    Journal Title

    Neuroscience

    Volume

    146

    Issue/Number

    2

    Publication Date

    1-1-2007

    Document Type

    Article

    Language

    English

    First Page

    481

    Last Page

    488

    WOS Identifier

    WOS:000246535000001

    ISSN

    0306-4522

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