High-throughput platform comprising microtissues perfused with living microvessels

Patent metadata

Field	Value
Publication number	US-9810685-B2
Application number	US-201113253820-A
Country	US
Kind code	B2
Filing date	Oct 5, 2011
Priority date	Oct 5, 2010
Publication date	Nov 7, 2017
Grant date	Nov 7, 2017

Abstract

Provided is a process for creating a 3D metabolically active microtissue perfused with living microvessels which have a direct fluidic connection with neighboring microfluidic channels. The process comprises preparing a template comprising a plurality of channels, and creating a network within said channels, said network comprising microfluidic channels, metabolically active living microvessels, and microtissues. The microvessels can sprout from said microvessels and/or form within the microtissue in response to a stimulus applied from said microfluidic channels or stimulus derived from the said tissues. In another embodiment, a device is provided comprising a supportive structure, one or more microfluidic channels, one or more microtissue compartments, and one or more microvessels, whereby the microvessels connect said microfludic channels and microtissue and perfuse the microtissue to deliver fluid from the microfluidic channels to the microtissues.

First claim

The invention claimed is:

1. A three-dimensional cell system, comprising:
a plurality of microfabricated microfluidic channels including a first microfabricated microfluidic channel and a second microfabricated microfluidic channel formed within a non-biological supportive material;
one or more microfabricated compartments formed within the non-biological supportive material;
cells combined with a biomimetic matrix and residing in at least one microfabricated compartment of the one or more microfabricated compartments; and
one or more living microvessels formed subsequent to placement of the cells combined with the biomimetic matrix within the at least one microfabricated compartment, wherein the one or more living microvessels including a first living microvessel that connects to the first microfabricated microfluidic channel and the second microfabricated microfluidic channel, the first living microvessel having a lumen and running from the first microfabricated microfluidic channel, through the at least one microfabricated compartment, and to the second microfabricated microfluidic channel, and the one or more living microvessels perfusing the cells combined with the biomimetic matrix and coupling together the plurality of microfabricated microfluidic channels,
wherein the one or more living microvessels allow for (i) delivery of nutrients from a fluid flowing from the first microfabricated microfluidic channel of the plurality of microfabricated microfluidic channels to the cells combined with the biomimetic matrix and (ii) removal of at least waste products from the fluid flowing to the second microfabricated microfluidic channel of the plurality of microfabricated microfluidic channels,
wherein the fluid flows inside the lumen of the one or more living microvessels, and wherein the one or more living microvessels are selected from a group consisting of (i) one or more living lymphatic vessels and (ii) one or more living blood vessels.

2. The three-dimensional cell system of claim 1 , wherein the plurality of microfabricated microfluidic channels, or a combination of the plurality of microfabricated microfluidic channels and the cells combined with the biomimetic matrix, is seeded with cells obtained from an individual.

3. The three-dimensional cell system of claim 1 , wherein the plurality of microfabricated microfluidic channels comprise seeded endothelial cells, seeded stromal cells, or both the seeded endothelial cells and the seeded stromal cells.

4. The three-dimensional cell system of claim 1 , wherein the cells combined with the biomimetic matrix comprise human cells, cancer cells, or patient-specific cells.

5. The three-dimensional cell system of claim 4 , wherein the human cells comprise normal or diseased cells selected from a group consisting of stem cells, endothelial cells, stromal cells, epithelial cells, neuronal cells, connective cells, myocardial cells, hepatocytes, renal, heart cells, liver cells, pancreatic cells, muscle cells, brain cells, kidney cells, tumor cells, and patient-specific cells.

6. The three-dimensional cell system of claim 1 , wherein the first living microvessel allows for delivery of the nutrients within the fluid that provides viability of the cells combined with the biomimetic matrix.

7. The three-dimensional cell system of claim 1 , wherein the fluid includes nutrients, and further includes a stimulant, a reagent, or a combination thereof, for testing a response of one or more of the cells combined with the biomimetic matrix.

8. The three-dimensional cell system of claim 7 , wherein the fluid includes a drug that corresponds to the stimulant, the reagent, or the combination of the stimulant and the reagent.

9. The three-dimensional cell system of claim 1 , wherein survival of the cells combined with the biomimetic matrix-depends, at least in part, on movement of the fluid through the first living microvessel.

10. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels sprout from endothelial cells inside the plurality of microfabricated microfluidic channels in response to a stimulus from within the cells combined with the biomimetic matrix.

11. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels sprout from endothelial cells inside the cells combined with the biomimetic matrix in response to a stimulus from within the cells combined with the biomimetic matrix.

12. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels sprout from endothelial cells inside the cells combined with the biomimetic matrix in response to a stimulus, the stimulus being a condition induced by mechanical, chemical or a combination of mechanical and chemical factors.

13. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels are formed in response to a stimulus, the stimulus being based, at least in part, on a controlled pressure within at least one of the plurality of microfabricated microfluidic channels.

14. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels are formed in response to a stimulus, the stimulus being based, at least in part, on a controlled pressure within the at least one microfabricated compartment.

15. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels comprises a microvessel network including a plurality of interconnected living microvessels.

16. The three-dimensional cell system of claim 1 , wherein the supportive material comprises polydimethyl siloxane (PDMS).

17. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels include a second living microvessel that connects to the first and second microfabricated microfluidic channels.

18. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels are the one or more living blood vessels.

19. The three-dimensional cell system of claim 18 , wherein the one or more living blood vessels are metabolically active.

20. The three-dimensional cell system of claim 18 , wherein the one or more living blood vessels comprise endothelial cells derived from endothelial cells seeding the plurality of microfabricated microfluidic channels.

21. The three-dimensional cell system of claim 18 , wherein the one or more living blood vessels comprise endothelial cells derived from the cells combined with the biomimetic matrix.

22. The three-dimensional cell system of claim 18 , wherein the one or more living blood vessels form developing blood vessel networks.

23. The three-dimensional cell system of claim 18 , wherein the one or more living blood vessels are capable of sprouting additional blood vessels.

24. The three-dimensional cell system of claim 18 , wherein the one or more living blood vessels are stimulated to form in response to fluid flow rate in the plurality of microfabricated microfluidic channels.

25. The three-dimensional cell system of claim 1 , wherein the one or more living microvessels are the one or more living lymphatic vessels.

Assignees

• George Steven C
• Hughes Christopher C W
• Lee Abraham P
• Moya Monica
• Hsu Yu-Hsiang
• Univ California

Inventors

• George Steven C
• Hughes Christopher C W
• Lee Abraham P
• Moya Monica
• Hsu Yu-Hsiang

Classifications

• C12N5/0062

  General methods for three-dimensional culture · CPC title

• C12N5/0691

  Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• C12N2535/00

  Supports or coatings for cell culture characterised by topography · CPC title

• G01N33/5082 Primary

  Supracellular entities, e.g. tissue, organisms · CPC title