Oscillating cell culture bioreactor

• US 9,217,129 B2
• Filed: 02/08/2008
• Issued: 12/22/2015
• Est. Priority Date: 02/09/2007
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Freed, Lisa E., Langer, Robert S., Moretti, Matteo G.
• Primary Examiner(s): Beisner, William H
• Application Number:
  US12/526,300
• Publication Number:
  US 20100297233A1
• Time in Patent Office: 2,874 Days
• Field of Search: 435/284.1, 435/293.1, 623/916, 623/917, 623/921
• US Class Current: 1/1
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/02 Form or structure of the vessel large containers B65D88/00
• CPC Class Codes: C12M 23/06 Tubular C12M23/08, C12M23/16 take precedence
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12M 27/10 Rotating vessel

Abstract

Methods and devices for cell or tissue culture are provided. One aspect provides a bioreactor having a gas permeable, closed-loop chamber for cell or tissue culture, and an oscillating means for moving the gas permeable, closed-loop chamber bidirectionally along an axis horizontal to an axis normal to the closed-loop chamber to force convection of cells and fluid in the gas permeable, closed-loop chamber. The bioreactor optionally includes a tissue engineering scaffold, an inlet means, an outlet means, and integrated sensors. Another aspect provides a bioreactor having a plurality of gas permeable, closed-loop chambers for cell or tissue culture. Methods of culturing cells and producing tissue constructs are also provided.

Claims

1. A bioreactor comprising

one or more gas permeable, closed-loop chambers which can contain fluid and gases and a tissue engineering scaffold for cell or tissue culture;

a tissue engineering scaffold or tissue engineering matrix secured directly to the wall of the closed-loop chamber or placed within a means for retaining the scaffold which is secured directly to the wall of closed-loop chamber so that fluid, cells and gases are forced through the entire thickness of the scaffold; and

an oscillator and controller for moving the gas permeable, closed-loop chamber bidirectionally along an axis horizontal to an axis normal to the closed-loop chamber to force cells and fluid within the gas permeable, closed-loop chamber by convective flow through the secured tissue engineering scaffold.

2. The bioreactor of claim 1, wherein the gas permeable, closed-loop chamber is toroidal.

3. The bioreactor of claim 1, further comprising a plurality of gas permeable, closed-loop chambers.

4. The bioreactor of claim 1, further comprising a housing for providing controlled environmental conditions.

5. The bioreactor of claim 4, comprising means for maintaining humidity, means for regulating temperature at about 35-37°

C., and means for controlling gases to about 21% oxygen and between about 5-10% CO₂.

6. The bioreactor of claim 4, wherein the housing oxygenates and buffers the pH of a fluid in the gas permeable, closed-loop chamber.

7. The bioreactor of claim 1, wherein the gas permeable, close-looped chamber comprises a gas-permeable polymer.

8. The bioreactor of claim 7, wherein the gas-permeable polymer is silicone rubber.

9. The bioreactor of claim 7 wherein the polymer is translucent or transparent.

10. The bioreactor of claim 1, further comprising therein cells on the scaffold or tissue engineering matrix.

11. The bioreactor of claim 10 wherein the cells are mammalian cells selected from the group consisting of cardiac cells, mesenchymal cells, fibroblasts, interstitial cells, neural cells, endothelial cells, smooth or skeletal muscle cells, myocytes, chrondocytes, adipocytes, fibromyoblasts, ectodermal cells, ductile and skin cells, hepatocytes, Islet cells, cells present in the intestine, parenchymal cells, osteoblasts, bone marrow cells, blood cells, and stem cells.

12. The bioreactor of claim 1, further comprising a hydrogel in combination with the scaffold or matrix.

13. The bioreactor of claim 1 wherein the scaffold is a hydrogel.

14. The bioreactor of claim 1 comprising cells to be cultured to form cardiac tissue, cartilage, bone, muscle or skin tissue.

15. The bioreactor of claim 1 wherein convective flow is achieved by trapping a bubble at the top of the closed-loop chamber or using a partially occlusive roller which compresses tubing at the top of the closed-loop chamber.

16. A method for culturing cells comprising

culturing the cells in a bioreactor comprising

one or more gas permeable, closed-loop chambers which can contain fluid and gases and a tissue engineering scaffold for cell or tissue culture;

a tissue engineering scaffold or tissue engineering matrix secured directly to the wall of the closed-loop chamber or placed within a means for retaining the scaffold which is secured directly to the wall of the closed-loop chamber so that fluid, cells and gases are forced through the entire thickness of the scaffold; and

an oscillator and controller for moving the gas permeable, closed-loop chamber bidirectionally along an axis horizontal to an axis normal to the closed-loop chamber to force cells and fluid within the gas permeable, closed-loop chamber by convective flow through the secured tissue engineering scaffold.

17. The method of claim 16 for producing a scaffold or tissue engineering matrix comprising

placing a scaffold or tissue engineering matrix and culture media in the gas permeable, closed-loop chamber;

sealing the gas permeable, closed-loop chamber;

oscillating the gas permeable, closed-loop chamber bidirectionally along an axis horizontal to an axis normal to the closed-loop chamber to force convection of the culture media and cells through the scaffold; and

culturing the cells in the bioreactor.

18. The method of claim 17, wherein the cells are selected from the group consisting of cardiac cells, mesenchymal cells, fibroblasts, interstitial cells, neural cells, endothelial cells, smooth or skeletal muscle cells, myocytes, chrondocytes, adipocytes, fibromyoblasts, ectodermal cells, ductile and skin cells, hepatocytes, Islet cells, cells present in the intestine, parenchymal cells, osteoblasts, bone marrow cells, blood cells, and stem cells.

19. The method of claim 16 wherein convective flow is achieved by trapping a bubble at the top of the closed-loop chamber or using a partially occlusive roller which compresses tubing at the top of the closed-loop chamber.