Muscle chips and methods of use thereof

Patent metadata

Field	Value
Publication number	US-9857356-B2
Application number	US-201214362287-A
Country	US
Kind code	B2
Filing date	Dec 10, 2012
Priority date	Dec 9, 2011
Publication date	Jan 2, 2018
Grant date	Jan 2, 2018

Abstract

The present invention provides high throughput assays for identifying compounds that modulate a contractile function, as well as devices suitable for these assays.

First claim

What is claimed:

1. A fluidic device for measuring a contractile function, the fluidic device comprising:
a chamber comprising a solid support structure;
a porous membrane dividing the chamber into a first channel and a second channel;
cells adhered to the porous membrane and disposed in the first channel of the chamber;
a plurality of muscle tissue strips disposed in the second channel of the chamber; and
a device to measure contractility of the plurality of muscle strips;
wherein the cells adhered to the porous membrane are selected from the group consisting of epithelial cells and endothelial cells; and
wherein the plurality of muscle tissue strips comprise a flexible polymer layer and/or a hydrogel layer and a population of isolated muscle cells seeded on the flexible polymer layer and/or the hydrogel layer in a predetermined pattern, wherein said muscle cells form a muscle tissue structure that can perform a contractile function.

2. The fluidic device of claim 1 , wherein the isolated muscle cells are selected from the group consisting of cardiac muscle cells, ventricular cardiac muscle cells, atrial cardiac muscle cells, striated muscle cells, smooth muscle cells, and vascular smooth muscle cells.

3. The fluidic device of claim 1 , further comprising:
a second chamber comprising a second solid support structure;
a second porous membrane dividing the second chamber into a first channel and a second channel;
cells adhered to the second porous membrane and disposed in the first channel of the second chamber; and
a monolayer of muscle cells and an electrophysiological capturing device disposed in the second channel of the second chamber, wherein said muscle cells form a muscle tissue structure that can perform a contractile function.

4. A fluidic device for measuring a contractile function, the fluidic device comprising:
a solid support structure comprising a first chamber and a second chamber operably connected, wherein said first chamber comprises a plurality of muscle tissue strips and a device to measure contractility,
wherein the plurality of muscle tissue strips comprise a flexible polymer layer and/or a hydrogel layer and a population of isolated diseased cells seeded on the flexible polymer layer and/or the hydrogel layer in a predetermined pattern, wherein said diseased muscle cells form a muscle tissue structure that can perform a contractile function; and
wherein said second chamber comprises a plurality of muscle tissue strips and a device to measure contractility, wherein the plurality of muscle tissue strips comprise a flexible polymer layer and/or a hydrogel layer and a population of isolated healthy muscle cells seeded on the flexible polymer layer and/or the hydrogel layer in a predetermined pattern, wherein said healthy muscle cells form a muscle tissue structure that can perform a contractile function.

5. A fluidic device for measuring a contractile function, the fluidic device comprising:
a first chamber comprising a solid support structure;
a plurality of muscle tissue strips disposed in the first chamber, wherein the plurality of muscle tissue strips comprise a flexible polymer layer and/or a hydrogel layer and a population of isolated airway smooth muscle cells seeded on the flexible polymer layer and/or the hydrogel layer in a predetermined pattern, wherein said airway smooth muscle cells form a muscle tissue structure that can perform a contractile function; and
a porous membrane having epithelial cells adhered thereto and exposed to air flow situated above the plurality of muscle tissue strips, wherein the porous membrane divides the first chamber into a first channel and a second channel with the epithelial cells disposed in the first channel and the plurality of muscle tissue strips disposed in the second channel.

6. The fluidic device of claim 5 , further comprising a second chamber operably connected to the first chamber,
wherein said population of isolated airway smooth muscle cells of said plurality of muscle tissue strips disposed in said first chamber comprises a population of isolated healthy airway smooth muscle cells; and,
said second chamber comprises a plurality of muscle tissue strips comprising a population of isolated diseased airway smooth muscle cells.

7. A method for identifying a compound that modulates a contractile function, the method comprising
providing the fluidic device of any one of claims 1 - 5 and 6 ; and
determining the effect of the test compound on a contractile function of said plurality of muscle tissue strips in the presence and absence of the test compound, wherein a modulation of the contractile function of said plurality of muscle tissue strips in the presence of said test compound as compared to the contractile function in the absence of said test compound indicates that said test compound modulates a contractile function, thereby identifying a compound that modulates a contractile function.

8. The method of claim 7 , wherein the contractile function is a biomechanical activity.

9. The method of claim 7 , wherein the contractile function is an electrophysiological activity.

10. The method of claim 9 , wherein the electrophysiological activity is a voltage parameter selected from the group consisting of action potential, action potential duration (APD), conduction velocity (CV), refractory period, wavelength, restitution, bradycardia, tachycardia, reentrant arrhythmia, intracellular calcium transient, transient amplitude, rise time as a measure of contraction, decay time as a measure of relaxation, total area under the transient as a measure of contractile force, spontaneous calcium release voltage, and focal calcium release voltage.

11. The method of claim 7 , wherein the plurality of muscle tissue strips are cultured in the presence of a fluorophor.

12. The method of claim 11 , wherein said fluorophor is a voltage-sensitive dye or an ion-sensitive dye.

13. The method of claim 12 , wherein said ion-sensitive dye is a calcium sensitive dye.

14. The method of claim 7 , wherein said plurality of muscle tissue strips comprise cardiomyocytes.

15. The method of claim 7 , wherein said plurality of muscle tissue strips comprises vascular smooth muscle cells or vascular endothelial cells.

16. The method of claim 7 , wherein said plurality of muscle tissue strips comprise smooth muscle cells or striated muscle cells.

17. A method for identifying a compound useful for treating a muscle disease, the method comprising
providing the fluidic device of any one of claims 1 - 5 and 6 ;
contacting said plurality of muscle tissue strips with a test compound; and
determining the effect of the test compound on a contractile function of said plurality of muscle tissue strips in the presence and absence of the test compound, wherein a modulation of the contractile function of said plurality of muscle tissue strips in the presence of said test compound as compared to the contractile function in the absence of said test compound indicates that said test compound modulates a contractile function, thereby identifying a compound useful for treating a muscle disease.

18. A fluidic device for measuring a contractile function, the device comprising:
a chamber comprising a solid support structure;
a porous membrane dividing the chamber into a first channel and a second channel;
cells adhered to the porous membrane and disposed in the first channel of the chamber; and
a monolayer of muscle cells and an electrophysiological capturing device disposed in the second channel of the chamber,
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Assignees

• Harvard College
• President And Fellows Of Havard College

Inventors

• Parker Kevin Kit
• Goss Josue Adrian
• Grosberg Anna
• Alford Patrick W
• Feinberg Adam W
• Agarwal Ashutosh
• Mccain Megan Laura
• Lind Johan Ulrik

Classifications

• G01N2500/04

  Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A) · CPC title

• G01N33/5008 Primary

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics · CPC title

• G01N2500/10

  involving cells · CPC title

• G01N33/5088 Primary

  of vertebrates · CPC title

• G01N33/4836

  using multielectrode arrays · CPC title