Journal
BIOMATERIALS
Volume 31, Issue 18, Pages 4880-4888Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2010.02.055
Keywords
In vitro test; Muscle; Nerve tissue engineering; Neural cell; Surface modification; Self assembly
Funding
- NIH [5R01-NS050452]
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Neuromuscular junction (NMJ) formation, occurring between motoneurons and skeletal muscle, is a complex multistep process involving a variety of signaling molecules and pathways. In vitro motoneuron-muscle co-cultures are powerful tools to study the role of different growth factors, hormones and cellular structures involved in NMJ formation. In this study, a serum-free culture system utilizing defined temporal growth factor application and a non-biological substrate resulted in the formation of robust NMJs. The system resulted in long-term survival of the co-culture and selective expression of neonatal myosin heavy chain, a marker of myotube maturation. NMJ formation was verified by colocalization of dense clusters of acetylcholine receptors visualized using alpha-bungarotoxin and synaptophysin containing vesicles present in motoneuron axonal terminals. This model will find applications in basic NMJ research and tissue engineering applications such as bio-hybrid device development for limb prosthesis and regenerative medicine as well as for high-throughput drug and toxin screening applications. (C) 2010 Elsevier Ltd. All rights reserved.
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