Bioreactor design and process for engineering tissue from cells

• US 6,979,308 B1
• Filed: 06/05/2000
• Issued: 12/27/2005
• Est. Priority Date: 06/03/1999
• Status: Expired due to Fees

• Current Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Original Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Inventors: Wolfe, Stephen P., MacDonald, Jeffrey M.
• Primary Examiner(s): Bianco, Patricia
• Assistant Examiner(s): DEAK, LESLIE R
• Application Number:
  US09/586,981
• Time in Patent Office: 2,031 Days
• Field of Search: 604/4.01, 604/5.01, 604/6.09, 604/6.01, 604/6.04, 604/6.14, 604/7, 604/23, 604/500, 604/522, 604/48, 435/407, 4352971- 4, 435/398, 435/400, 435/366, 435/370, 435/373, 435/374, 435/283.1, 435/289.1, 435/294.1, 435/401, 422/44, 422/45, 422/48, 21032187- 89, 210/321.6, 210/500.1, 210/500.21, 210/321.64, 210/294, 210/295, 210/314, 210/316, 210/321.84, 210/500.23, 210/600, 210/634, 210/641, 210/644, 210/645, 210/255, 210/194, 210/195.1, 210/195.2, 210/198.1, 210/200, 210/207, 210/203, 210/252, 2102571- 2
• US Class Current: 604/6.09
• CPC Class Codes: B01D 63/02 Hollow fibre modules
• CPC Class Codes: B01D 63/0222 using centrifugal forces
• CPC Class Codes: B01D 63/0233 forming the bundle
• CPC Class Codes: B01D 63/026 Wafer type modules or flat-surface type modules
• CPC Class Codes: C12M 23/34 Internal compartments or partitions
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12M 29/16 Hollow fibers hollow fiber modules in general B01D63/02

Abstract

A scaled-up multi-coaxial fiber bioreactor, and variations of this bioreactor. The device is characterized by a hollow housing and an array of from about 20 to about 400 modules of hollow fibers, where each module includes at least three coaxial semipermeable hollow fibers. The innermost fiber provides a boundary for an innermost compartment which is connected to inlet and outlet ports. Arranged coaxially around the central hollow fiber are several other hollow fibers with their respective compartments, each compartment defined by a respective annular space between adjacent fibers and each including inlet and outlet ports. An outermost compartment for permitting integral aeration is the space between the outer side of the outermost fibers and the inner side of the housing, and has inlet and outlet ports. The hollow housing has inlet and outlet manifolds and flow distributors for each of the compartments. In a preferred embodiment the bioreactor is used as an extracorporeal liver. Liver cells, are introduced into one or more annular compartments and media and aeration are provided in others. Plasma from an ailing patient is introduced into another compartment for biotransformation of blood-borne toxins and biosynthesis of proteins, lipids, and other metabolic products.

Claims

1. A bioreactor, comprising:

(a) a housing having an inner side comprising;

a gas introduction means integral to the housing; and

a gas expiration means integral to the housing;

(b) a plurality of modules of hollow fibers, residing within the housing, each module comprising;

(i) at least three coaxially arranged hollow fibers, each hollow fiber having an inner side and an outer side, including an innermost hollow fiber and an outermost hollow fiber;

(ii) at least three compartments, comprising;

a first compartment defined by an inner side of an innermost hollow fiber and at least two compartments defined by a respective annular space between adjacent fibers of the at least three coaxially arranged hollow fibers; and

(c) an outermost compartment defined by a space within the inner side of the housing which is not occupied by the plurality of modules;

in which at least one compartment is charged with a gaseous nutrient substantially free of non-gaseous fluids.

2. The bioreactor of claim 1, where the hollow fibers are semipermeable.

3. The bioreactor of claim 2, where the hollow fibers comprise a material selected from the group consisting of polysulfone, polypropylene, nylon, polyester, polytetrafluoroethylene, cellulose acetate, and mixed esters of cellulose.

4. The bioreactor of claim 1, where the first compartment, the at least one additional compartment and the outermost compartment each further comprise at least one inlet port and at least one outlet port.

5. The bioreactor of claim 4, where the housing further comprises at least one inlet manifold and at least one outlet manifold for the first compartment and at least one inlet manifold and at least one outlet manifold for each additional compartment.

6. The bioreactor of claim 5, further comprising:

microfibers substantially parallel to the modules of hollow fibers.

7. The bioreactor of claim 6, where the microfibers further comprise at least one aeration inlet port and at least one aeration outlet port.

8. The bioreactor of claim 5, where the housing has a first end and a second end, and

where each inlet port and each exit port are at the first end of the housing.

9. The bioreactor of claim 5, where at least one manifold further comprises a flow distributor.

10. The bioreactor of claim 9, where at least one compartment further comprises an extracellular matrix.

11. The bioreactor of claim 1, where the bioreactor further comprises at least 10⁹ cells.

12. The bioreactor of claim 11, where the cells are liver cells.

13. The bioreactor of claim 12, where the liver cells are selected from the group consisting of porcine liver cells and human liver cells.

14. A method of supplying cell biosynthesis products to a patient in need thereof, comprising:

pumping intravenous feeding solution through a compartment of the bioreactor of claim 5;

collecting the output; and

intravenously feeding the output to the patient.

15. The bioreactor of claim 1, where at least one annular space is about 0.2 millimeters to about 0.8 millimeters.

16. The bioreactor of claim 1, where the bioreactor is sterilized by a means selected from the group consisting of autoclaving, ethylene oxide and gamma radiation.

17. The bioreactor of claim 1, wherein the innermost hollow fiber has a length of about 2 centimeters to about 50 centimeters.

18. The bioreactor of claim 1, where at least one coaxial hollow fiber is saturated with perfluorocarbon.

19. The bioreactor of claim 1, where at least one coaxial hollow fiber has a pore size less than 1×

10<sup>−

6</sup> m.

20. The bioreactor of claim 1, where at least one coaxial hollow fiber has a pore size less than 0.05×

10<sup>−

6</sup> m.

21. The bioreactor of claim 1, where at least one coaxial hollow fiber has a pore size less than 0.1×

10⁶ m.

22. The bioreactor of claim 1, where at least one compartment further comprises cells mixed with an extracellular matrix.

23. The bioreactor of claim 1 in which the gaseous nutrient includes oxygen.

24. The bioreactor of claim 23 in which the gaseous nutrient further includes carbon dioxide.

25. The bioreactor of claim 1 in which the gaseous nutrient includes air.

26. A serially-linked bioreactor, comprising a plurality of bioreactor subunits, each bioreactor subunit comprising:

(a) a housing having an inner side comprising;

a gas introduction means integral to the housing; and

a gas expiration means integral to the housing;

(b) a plurality of modules of hollow fibers, residing within the housing, each module comprising;

(i) at least three coaxially arranged hollow fibers, each hollow fiber having an inner side and an outer side, including an innermost hollow fiber and an outermost hollow fiber;

(ii) at least three compartments, comprising;

a first compartment defined by an inner side of an innermost hollow fiber; and

at least two compartments defined by a respective annular space between adjacent fibers of the at least three coaxially arranged hollow fibers; and

(c) an outermost compartment defined by a space within the inner side of the housing which is not occupied by the plurality of modules; and

(d) at least one compartment of one bioreactor subunit linked serially to at least one compartment of at least one other bioreactor subunit;

in which at least one compartment is charged with a gaseous nutrient substantially free of non-gaseous fluids.

27. The bioreactor of claim 26, where each bioreactor subunit further comprises at least 10⁹ cells.

28. The bioreactor of claim 27, where the cells are liver cells.

29. The bioreactor of claim 28 where the cells are selected from the group consisting of human liver cells and porcine liver cells.

30. The bioreactor of claim 27, where at least one compartment of each bioreactor subunit further comprises an extracellular matrix.

31. A method of treating a patient in need thereof comprising:

(a) introducing plasma of a patient into a bioreactor subunit of the serially linked bioreactor of claim 26,

(b) forcing at least a portion of the plasma to flow radially through a cell compartment of the bioreactor subunit to form a biotransformed effluent;

(c) introducing the biotransformed effluent into a second bioreactor subunit of the bioreactor of claim 26;

(d) forcing at least a portion of the biotransformed effluent to flow radially through a cell compartment of the second bioreactor subunit to form supplemented plasma; and

(e) returning the supplemented plasma to the patient'"'"'s circulatory system.

32. The method of claim 31 in which the gaseous nutrient includes oxygen.

33. The method of claim 32 in which the gaseous nutrient further includes carbon dioxide.

34. The method of claim 31 in which the gaseous nutrient includes air.

35. The serially linked bioreactor of claim 26 in which the gaseous nutrient includes oxygen.

36. The serially linked bioreactor of claim 35 in which the gaseous nutrient further includes carbon dioxide.

37. The serially linked bioreactor of claim 26 in which the gaseous nutrient includes air.