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Saini J, Faroni A, Abd Al Samid M, et al. Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells. Stem Cells Cloning. 2019;12:1-9doi: 10.2147/SCCAA.S187655.
Saini, J., Faroni, A., Abd Al Samid, M., Reid, A. J., Lightfoot, A. P., Mamchaoui, K., Mouly, V., Butler-Browne, G., McPhee, J. S., Degens, H., & Al-Shanti, N. (2019). Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells. Stem cells and cloning : advances and applications, 121-9. https://doi.org/10.2147/SCCAA.S187655
Saini, Jasdeep, et al. "Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells." Stem cells and cloning : advances and applications vol. 12 (2019): 1-9. doi: https://doi.org/10.2147/SCCAA.S187655
Saini J, Faroni A, Abd Al Samid M, Reid AJ, Lightfoot AP, Mamchaoui K, Mouly V, Butler-Browne G, McPhee JS, Degens H, Al-Shanti N. Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells. Stem Cells Cloning. 2019 Feb 20;12:1-9. doi: 10.2147/SCCAA.S187655. eCollection 2019. PMID: 30863121; PMCID: PMC6388735.
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Stem Cells Cloning. 2019 Feb 20;12:1-9. doi: 10.2147/SCCAA.S187655. eCollection 2019.

Simplified in vitro engineering of neuromuscular junctions between rat embryonic motoneurons and immortalized human skeletal muscle cells.

Stem cells and cloning : advances and applications

Jasdeep Saini, Alessandro Faroni, Marwah Abd Al Samid, Adam J Reid, Adam P Lightfoot, Kamel Mamchaoui, Vincent Mouly, Gillian Butler-Browne, Jamie S McPhee, Hans Degens, Nasser Al-Shanti

Affiliations

  1. Musculoskeletal Science & Sports Medicine Research Centre, School of Healthcare Science, Manchester Metropolitan University, Manchester, UK, [email protected].
  2. Blond McIndoe Laboratories, Division of Cell Matrix Biology and Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Manchester Academic Health Science Centre, Manchester, UK.
  3. Department of Plastic Surgery & Burns, University Hospitals of South Manchester, Manchester Academic Health Science Centre, Manchester, UK.
  4. Center for Research in Myology, Sorbonne Université- INSERM, Paris, France.
  5. Department of Sport and Exercise Science, Manchester Metropolitan University, Manchester, UK.
  6. Institute of Sport Science and Innovations, Lithuanian Sports University, Kaunas, Lithuania.
  7. University of Medicine and Pharmacy of Targu Mures, Targu Mures, Romania.

PMID: 30863121 PMCID: PMC6388735 DOI: 10.2147/SCCAA.S187655

Abstract

BACKGROUND: Neuromuscular junctions (NMJs) consist of the presynaptic cholinergic motoneuron terminals and the corresponding postsynaptic motor endplates on skeletal muscle fibers. At the NMJ the action potential of the neuron leads, via release of acetylcholine, to muscle membrane depolarization that in turn is translated into muscle contraction and physical movement. Despite the fact that substantial NMJ research has been performed, the potential of in vivo NMJ investigations is inadequate and difficult to employ. A simple and reproducible in vitro NMJ model may provide a robust means to study the impact of neurotrophic factors, growth factors, and hormones on NMJ formation, structure, and function.

METHODS: This report characterizes a novel in vitro NMJ model utilizing immortalized human skeletal muscle stem cells seeded on 35 mm glass-bottom dishes, cocultured and innervated with spinal cord explants from rat embryos at ED 13.5. The cocultures were fixed and stained on day 14 for analysis and assessment of NMJ formation and development.

RESULTS: This unique serum- and trophic factor-free system permits the growth of cholinergic motoneurons, the formation of mature NMJs, and the development of highly differentiated contractile myotubes, which exhibit appropriate configuration of transversal triads, representative of in vivo conditions.

CONCLUSION: This coculture system provides a tool to study vital features of NMJ formation, regulation, maintenance, and repair, as well as a model platform to explore neuromuscular diseases and disorders affecting NMJs.

Keywords: NMJ; coculture; motoneuron; motor neuron; myoblast; myotube; neuromuscular junction

Conflict of interest statement

Disclosure The authors report no conflicts of interest in this work.

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