A customizable microfluidic platform for medium-throughput modeling of neuromuscular circuits

Biomaterials. 2019 Dec;225:119537. doi: 10.1016/j.biomaterials.2019.119537. Epub 2019 Oct 8.

Abstract

Neuromuscular circuits (NMCs) are vital for voluntary movement, and effective models of NMCs are needed to understand the pathogenesis of, as well as to identify effective treatments for, multiple diseases, including Duchenne's muscular dystrophy and amyotrophic lateral sclerosis. Microfluidics are ideal for recapitulating the central and peripheral compartments of NMCs, but myotubes often detach before functional NMCs are formed. In addition, microfluidic systems are often limited to a single experimental unit, which significantly limits their application in disease modeling and drug discovery. Here, we developed a microfluidic platform (MFP) containing over 100 experimental units, making it suitable for medium-throughput applications. To overcome detachment, we incorporated a reactive polymer surface allowing customization of the environment to culture different cell types. Using this approach, we identified conditions that enable long-term co-culture of human motor neurons and myotubes differentiated from human induced pluripotent stem cells inside our MFP. Optogenetics demonstrated the formation of functional NMCs. Furthermore, we developed a novel application of the rabies tracing assay to efficiently identify NMCs in our MFP. Therefore, our MFP enables large-scale generation and quantification of functional NMCs for disease modeling and pharmacological drug targeting.

Keywords: Microfluidics; Motor unit; Neuromuscular circuit; Rabies viral tracing; Skeletal muscle; poly(ethylene-alt-maleic anhydride).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion / drug effects
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Dimethylpolysiloxanes / chemistry
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Laminin / pharmacology
  • Maleates / chemistry
  • Microfluidics / methods*
  • Motor Neurons / cytology
  • Motor Neurons / drug effects
  • Muscle Fibers, Skeletal / cytology
  • Muscle Fibers, Skeletal / drug effects
  • Neuromuscular Junction / physiology*
  • Peptides / pharmacology
  • Plasma Gases / chemistry
  • Polyethylenes / chemistry

Substances

  • Dimethylpolysiloxanes
  • Laminin
  • Maleates
  • Peptides
  • Plasma Gases
  • Polyethylenes
  • polyornithine
  • baysilon
  • ethylene-maleic anhydride copolymer