Perfused three-dimensional cell/tissue disease models

• US 20050260745A1
• Filed: 05/19/2005
• Published: 11/24/2005
• Est. Priority Date: 05/19/2004
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology, University of Pittsburgh of The Commonwealth System of Higher Education
• Original Assignee: Commonwealth of Massachusetts, Commonwealth Of Pennsylvania, Massachusetts Institute of Technology, University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Inventors: Domansky, Karel, Wells, Alan, Griffith, Linda G., Tannenbaum, Steven R.
• Granted Patent: US 8,318,479 B2
• Time in Patent Office: Days
• US Class Current: 435/294.100
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0638 membrane valves, flap valves
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/40 Manifolds; Distribution pieces fluid transfer means B01L3/563
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12M 29/10 Perfusion

Abstract

A system has been constructed that recapitulate the features of a capillary bed through normal human tissue. The system facilitates perfusion of three-dimensional (3D) cell monocultures and heterotypic cell co-cultures at the length scale of the capillary bed. A major feature is that the system can be utilized within a “multiwell plate” format amenable to high-throughput assays compatible with the type of robotics commonly used in pharmaceutical development. The system provides a means to conduct assays for toxicology and metabolism and as a model for human diseases such as hepatic diseases, including hepatitis, exposure-related pathologies, and cancer. Cancer applications include primary liver cancer as well as metastases. The system can also be used as a means of testing gene therapy approaches for treating disease and inborn genetic defects.

Claims

1. An apparatus comprising

A dense array of perfusion bioreactor and reservoir pairs for cell or tissue culture, and

valves and pumps actuated in parallel via common control channels and re-circulating medium through the array of bioreactor and reservoir pairs.

2. The apparatus of claim 1 wherein each bioreactor comprises a well comprising a three-dimensional cell/tissue support structure.

3. The apparatus of claim 1 wherein the cell scaffold or carrier is made out of porous material.

4. The apparatus of claim 2 wherein the cell scaffold is formed by an array of microchannels in a solid film or sheet supported by a microporous filter or membrane.

5. The apparatus of claim 2 wherein the scaffolds can be ejected from the bioreactor wells.

6. The apparatus of claim 1 wherein each bioreactor of the array comprises a bioreactor well and its own reservoir well.

7. The apparatus of claim 6 wherein the bioreactor wells and reservoir wells are connected by fluidic channels allowing re-circulation of cell culture medium.

8. The apparatus of claim 1 wherein all bioreactors/reservoir pairs in the array are covered by a common removable lid.

9. The apparatus of claim 1 comprising a device for manual or robotic pipetting of cell/tissue seeding, agent addition, or sample collection.

10. The apparatus of claim 1 wherein each bioreactor/reservoir pair is fluidically isolated from all other bioreactor/reservoir pairs in the array.

11. The apparatus of claim 1 wherein the valves and pumps of all bioreactors in the array are actuated in parallel via common hydraulic or pneumatic control channels.

12. The apparatus of claim 1 wherein the bioreactor/reservoir pairs are fabricated or microfabricated in the fluidic manifold.

13. The apparatus of claim 1 wherein the control channels are fabricated or microfabricated in the control manifold.

14. The apparatus of claim 1 wherein diaphragm valves are created by sandwiching a monolithic elastomeric membrane between fluidic and control manifolds.

15. The apparatus of claim 14 wherein the membrane between the control and fluidic channels can be deflected by hydraulic or pneumatic actuation applied through the control channels.

16. The apparatus of claim 1 wherein cell culture medium in multiple bioreactors is pumped by sequential actuation of the valves connected in series.

17. The apparatus of claim 1 further comprising a first set of cells.

18. The apparatus of claim 17 further comprising a second set of cells introduced after the initial bioreactor tissue is established.

19. The apparatus of claim 18 wherein the second set of cells are tumor cells.

20. The apparatus of claim 18 where the second set of cells are stem cells.

21. The apparatus of claim 17 comprising tumor cells that grow in the environment of liver tissue.

22. The apparatus of claim 17 comprising tumor cells that can be studied for tumor cell behaviors.

23. The apparatus of claim 18 wherein molecular events can be visualized during tumor cell growth.

24. The apparatus of claim 1 wherein the first set of cells can be modified prior to or subsequent to introduction into the apparatus.

25. The apparatus of claim 1 comprising primary tumor cells from patients for diagnostic and prognostic testing.

26. The apparatus of claim 25 wherein the tumor cells can be assessed for sensitivity to an agent or gene therapy.

27. The apparatus of claim 21 wherein tumor cell sensitivity to an agent or gene therapy is linked to liver metabolism of set agent or gene therapy.

28. The apparatus of claim 20 wherein the second set of cells introduced are stem or progenitor cells and the stem or progenitor cells are induced to differentiate by the mature tissue.

29. The apparatus of claim 17 comprising mature cells inducible to replicate by manipulation of the flow rates or medium components in the system.