THREE DIMENSIONAL TISSUES FOR HIGH-THROUGHPUT ASSAYS

• US 20110118143A1
• Filed: 06/05/2009
• Published: 05/19/2011
• Est. Priority Date: 06/05/2008
• Status: Active Grant

• Current Assignee: InvivoSciences Inc.
• Original Assignee: InvivoSciences Inc.
• Inventors: Wakatsuki, Tetsuro
• Granted Patent: US 9,631,169 B2
• Time in Patent Office: Days
• US Class Current: 506/10
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: C40B 30/06 by measuring effects on living organisms, tissues or cells
• CPC Class Codes: G01N 33/5082 Supracellular entities, e.g. tissue, organisms

Abstract

Provided herein are methods of detecting responses of bio-artificial tissues to agents by performing assays using three-dimensional bio-artificial tissues. The methods are adaptable to high-throughput platforms.

Claims

1. A method of detecting the response of a tissue to an agent comprising:

(a) contacting a bio-artificial tissue with an agent, the bio-artificial tissue comprising cells and extracellular matrix, the bio-artificial tissue formed on a scaffold support without a fastener to facilitate tissue adhesion, the scaffold support being positioned above the bottom of a well,

(b) performing an assay using the bio-artificial tissue, wherein the assay produces an indicator; and

(c) detecting a level of the indicator in the well, wherein the level of the indicator is indicative of the response of the bio-artificial tissue to the agent.

2. The method of claim 1, wherein the cells are selected from the group consisting of a muscle cell, a non-muscle cell, an endothelial cell, and a cardiac cell.

3. The method of claim 1, wherein the well is positioned within a multi-well plate.

4. The method of claim 3, wherein the multi-well plate comprises from 2 to 10,000 wells.

5. The method of claim 1, wherein the scaffold support is a wire frame.

6. The method of claim 5, wherein the wire is stainless steel wire.

7. The method of claim 1, wherein the cells comprise cells known to be involved in a disease.

8. The method of claim 1, wherein the indicator is a colorimetric or a fluorometric indicator.

9. The method of claim 1, wherein the level of the indicator is detected using a microscope.

10. The method of claim 1, wherein the indicator is a radioactive label.

11. The method of claim 10, wherein the level of the indicator is detected using a scintillator.

12. The method of claim 1, wherein the level of the indicator is detected using a multi-well plate reader.

13. The method of claim 1, wherein the assay is selected from the group consisting of cell proliferation assays, cell death assays, apoptosis assays, protein expression assays, gene expression assays, enzymatic assays, and cell signaling assays.

14. The method of claim 1, wherein the agent is a candidate pharmaceutical.

15. The method of claim 1, wherein the method is a high throughput screening method.