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Engineering and Characterization of an Optogenetic Model of the Human Neuromuscular Junction
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Bioengineering
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JoVE Journal Bioengineering
Engineering and Characterization of an Optogenetic Model of the Human Neuromuscular Junction

Engineering and Characterization of an Optogenetic Model of the Human Neuromuscular Junction

DOI:
10.3791/63759-v

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11:07 min

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April 14, 2022

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, , , ,
1Department of Biomedical Engineering,Columbia University, 2Department of Medicine,Columbia University, 3Gladstone Institutes, 4College of Dental Medicine,Columbia University

Chapters

  • 00:04Introduction
  • 00:48Fabrication of Bioreactors
  • 02:31Skeletal Muscle Tissue Seeding (Day 3)
  • 03:22Motoneuron Differentiation (Days 0 – 14)
  • 04:01Seeding Motoneuron Aggregates in Bioreactor (Day 14)
  • 05:31Simultaneous Optical Stimulation and Video Recording of Neuromuscular Junction (NMJ) Function (Day 24+)
  • 07:36Batch Processing and Analysis (Day 24+)
  • 08:34Results: Stimulation, Recording, and Analysis of NMJ Function
  • 10:31Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a reproducible, automated, and unbiased imaging system for characterizing neuromuscular junction function using human engineered skeletal muscle tissue and optogenetic motoneurons. This system allows for the functional quantification of neuromuscular connectivity over time and detects diminished neuromuscular function caused by neurotoxins and myasthenia gravis patient serum.

Tags

Keywords: OptogeneticNeuromuscular JunctionNeuromuscular PathologiesTherapeuticsVideo ProcessingNeurotoxinsMyasthenia GravisPDMSCollagen GelMyoblastsMotoneuron Differentiation

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