Devices and methods comprising neuromuscular junctions

Patent metadata

Field	Value
Publication number	US-10935541-B2
Application number	US-201514821675-A
Country	US
Kind code	B2
Filing date	Aug 7, 2015
Priority date	Aug 7, 2014
Publication date	Mar 2, 2021
Grant date	Mar 2, 2021

Abstract

Disclosed herein are devices comprising one or more cantilevers comprising one or more neuromuscular junctions formed by a co-culture of myotubes and motoneurons. Disclosed herein are methods of using the disclosed devices comprising one or more cantilevers. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.

First claim

What is claimed is:

1. A device, comprising:
a first chamber comprising a myotube culture attached to a plurality of cantilevers;
a second chamber spaced from the first chamber by a barrier, the second chamber comprising a motoneuron culture attached to and in electrical communication with a microelectrode array, and
an automated detection system configured to detect a deflection of the at least one cantilevers, and
at least one motoneuron selected from the motoneuron culture, the at least one motoneuron comprising a cell body, an axon, and a terminal,
wherein the cell body is in electrical communication with the microelectrode array in the second chamber, the axon extends through an opening of the barrier, and the terminal is in electrophysiological communication with a myotube in the first chamber to form a neuromuscular junction, and
wherein electrical stimulation of the motoneuron culture via the microelectrode array causes electrophysiological signal transmission through the axon and the neuromuscular junction to a myotube of the myotube culture, causing the myotube to contract and deflect at least one cantilever of the plurality of cantilevers.

2. The device of claim 1 , wherein the automated detection system comprises a laser and a photo-detector.

3. The device of claim 1 , wherein muscle cells or myoblasts fuse together to form one or more of the myotubes.

4. The device of claim 1 , wherein the at least one cantilever comprises one or more piezoelectric materials.

5. The device of claim 4 , wherein the automated detection system comprises a transducer for detecting a change in electrical conductivity of the at least one cantilever.

6. The device of claim 4 , wherein piezoelectric materials comprise at least one of quartz, bone, sodium tungstate, zinc oxide, lead zirconate titanate, or a combination thereof.

7. The device of claim 1 , wherein the at least one cantilever comprises at least two cantilevers.

8. The device of claim 2 , wherein the automated detection system further comprises a plurality of linear actuators attached to XY translational stages that control the position of the laser and the photo-detector.

9. The device of claim 2 , further comprising a digitizer and a computer, wherein the photo-detector is in communication with the digitizer and wherein the digitizer is in communication with the computer.

10. The device of claim 1 , further comprising a temperature-controlled stage, wherein the at least one cantilever is maintained on the temperature-controlled stage, and wherein the temperature-controlled stage comprises one or more electrodes and one or more pulse generators.

11. The device of claim 10 , further comprising a digitizer and a computer, wherein the one or more pulse generators is in communication with the digitizer, and wherein the digitizer is in communication with the computer.

12. The device of claim 1 , wherein the at least one cantilever is surface-modified or surface-coated.

13. The device of claim 12 , wherein the at least one cantilever comprises a (3-Trimethoxysilyl propyl) diethylenetriamine (DETA) surface modification.

14. The device of claim 1 , wherein the myotubes are human or rat myotubes, and the motoneurons are human or rat motoneurons.

15. The device of claim 1 , wherein at least one of the myotubes or motoneurons is obtained from a subject diagnosed with or suspected of having a muscle wasting condition, a peripheral neuropathy, or both.

16. The device of claim 1 , further comprising a serum-free medium, wherein the co-culture is maintained in the serum-free medium.

17. The device of claim 16 , wherein the serum-free medium comprises neurobasal medium, B27, Glutamax, glial-derived neurotrophic factor, brain-derived neurotrophic factor, ciliary neurotrophic factor, insulin-like growth factor-1, neurotrophin-3, neurotrophin-4, mouse laminin, and cAMP.

18. The device of claim 1 , further comprising extracellular matrix molecules, chemotactic factors, or a combination thereof patterned on a surface of the device positioned between the first chamber and the second chamber.

19. The device of claim 1 , wherein the device is an in vitro device.

20. The device of claim 1 , further comprising a barrier arranged between the first chamber and the second chamber, the barrier comprising a plurality of openings.

21. The device of claim 20 , wherein the barrier further comprises a plurality of channels, and the axon extends through a channel of the plurality of channels.

Assignees

• Univ Central Florida Res Found Inc

Inventors

• Hickman James
• Smith Alexander
• Long Christopher
• Pirozzi Kristen

Classifications

• G01N33/5058

  Neurological cells · CPC title

• G01N33/48728

  Investigating individual cells, e.g. by patch clamp, voltage clamp (investigating individual particles in general G01N15/10) · CPC title

• G01N33/5061 Primary

  Muscle cells · CPC title