Biological artificial liver

• US 5,270,192 A
• Filed: 02/07/1991
• Issued: 12/14/1993
• Est. Priority Date: 02/07/1991
• Status: Expired due to Term

• Current Assignee: Monsanto Company (Bayer AG)
• Original Assignee: Monsanto Company (Bayer AG)
• Inventors: Li, Albert Pak-Hung, Whitehead, Timothy D., Barker, George E., Beck, Dale J.
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): Beisner, William H.
• Application Number:
  US07/832,461
• Time in Patent Office: 1,041 Days
• Field of Search: 435/1, 435/240.1, 435/240.23, 435/283, 435/171, 435/176, 435/284, 435/286, 435/287, 435/299, 435/310, 435/311, 604/4-6, 623/66
• US Class Current: 435/174
• CPC Class Codes: A61M 1/3472 with treatment of the filtrate
• CPC Class Codes: A61M 1/3489 by biological cells, e.g. bioreactor
• CPC Class Codes: A61M 1/3603 in the same direction
• CPC Class Codes: A61M 2202/0064 changed by biological action
• CPC Class Codes: A61M 2202/0415 Plasma
• CPC Class Codes: C12M 25/16 Particles; Beads; Granular material; Encapsulation chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12N 5/0671 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

Disclosed is a hepatocyte bioreactor or bioartificial liver which comprises a containment vessel having a perfusion inlet and a perfusion outlet; a matrix within the containment vessel so as to entrap hepatocyte aggregates within the containment vessel while allowing perfusion of the matrix. The invention is useful as an artificial liver.

Claims

1. A hepatocyte bioreactor, which comprises:

a containment vessel having an inlet which allows hepatocytes or aggregates thereof to ingress into said containment vessel and an outlet which allows hepatocytes or aggregates thereof to egress;

a matrix within said containment vessel into which said hepatocytes or aggregates thereof can ingress, said matrix being in fluid communication with said inlet and said outlet; and

a plurality of hepatocyte aggregates entrapped within said matrix wherein said hepatocyte aggregates are interconnected within said matrix by cell-cell contact.

2. The hepatocyte bioreaction of claim 1 wherein said hepatocyte aggregates are spheroids.

3. The hepatocyte bioreactor of claim 1 wherein said matrix is glass beads.

4. The hepatocyte bioreactor of claim 1 wherein said hepatocyte aggregates are essentially free of connective tissue.

5. The hepatocyte bioreactor of claim 1 wherein said matrix has a hepatocytes density of at least about 3.00×

10⁶ cells per ml of matrix.

6. A method for making a bioreactor, which comprises the steps of:

providing a containment vessel having an inlet which allows hepatocytes or aggregates thereof to ingress into said containment vessel and an outlet which allows hepatocytes or aggregates thereof to egress;

providing a matrix within said containment vessel into which said hepatocytes or aggregates thereof can ingress, said matrix being in fluid communication with said inlet and said outlet; and

entrapping a plurality of hepatocyte aggregates within said matrix wherein said hepatocyte aggregates are interconnected within said matrix by cell-cell contact.

7. The method for making a bioreactor as recited in claim 6 wherein said hepatocyte aggregates are spheroids.

8. The method for making a hepatocyte bioreactor of claim 6 wherein said matrix is glass beads.

9. The method for making a hepatocyte bioreactor of claim 6 wherein said hepatocyte aggregates are essentially free of connective tissue.

10. The method for making a heptocyte bioreactor of claim 6 wherein said matrix has a hepatocytes density of at least about 3.00×

10⁶ cells per ml of matrix.

11. The method for making a hepatocyte bioreactor of claim 6 wherein said hepatocyte aggregates are formed by perfusion of said hepatocytes through said matrix.

12. A method for using a hepatocyte bioreactor, which comprises the steps of:

providing a containment vessel having an inlet which allows hepatocytes or aggregates thereof to ingress into said containment vessel and an outlet which allows hepatocytes or aggregates thereof to egress;

providing a matrix within said containment vessel into which said hepatocytes or aggregates thereof can ingress, said matrix being in fluid communication with said inlet and said outlet;

entrapping a plurality of hepatocyte aggregates within said containment vessel wherein said hepatocyte aggregates are interconnected within said matrix by cell-cell contact; and

causing medium to be perfused through said inlet, said containment vessel and said outlet.

13. The method of using a hepatocyte bioreactor as recited in claim 12 wherein said plurality of hepatocyte aggregates are spheroids.

14. The method of using a hepatocyte bioreactor of claim 12 wherein said hepatocyte aggregates are formed by perfusion of said hepatocytes through said matrix.

15. The method of using a hepatocyte bioreactor as recited in claim 12 wherein said medium contains metabolites.

16. The hepatocyte bioreactor of claim 12 wherein said matrix is glass beads.

17. The method of using a hepatocyte bioreactor as recited in claim 12 wherein said hepatocyte aggregates are essentially free of connective tissue.

18. The method of using a hepatocyte bioreactor as recited in claim 12 wherein said matrix has a hepatocytes density of at least about 3.00×

10⁶ cells per ml of matrix.

19. A hepatocyte bioreactor liver assist system for treating a mammal in need of liver assistance, which comprises;

means for extracting blood from the mammal in need of liver assistance;

means for separating the blood of the mammal which is to be treated into cellular and non-cellular components;

a hepatocyte bioreactor comprising;

a containment vessel having an inlet which allows hepatocytes or aggregates thereof to ingress into said containment vessel and an outlet which allows hepatocytes or aggregates thereof to egress;

a matrix within said containment vessel into which said hepatocytes or aggregates thereof can ingress;

said matrix being in fluid communication with said inlet and aid outlet; and

a plurality of hepatocyte aggregates entrapped within said matrix wherein said hepatocyte aggregates are interconnected within said matrix by cell-cell contact;

means causing said noncellular blood components of the mammal which is to be treated to be perfused through said inlet, said matrix and said outlet; and

means to return said treated noncellular blood components and cellular blood components to said mammal.

20. The hepatocyte bioreactor liver assist system of claim 19 further comprising a means for maintaining a flow rate of medium through the hepatocyte bioreactor which is independent of the mammal.

21. The hepatocyte bioreactor liver assist system of claim 19 wherein said hepatocyte aggregates are spheroids.

22. The hepatocyte bioreactor liver assist system of claim 19 wherein said matrix is glass beads.

23. The hepatocyte bioreactor liver assist system of claim 19 wherein said hepatocyte aggregates are essentially free of connective tissue.

24. The hepatocyte bioreactor liver assist system of claim 19 wherein said matrix has a hepatocytes density of at least about 3.00×

10⁶ cells per ml of matrix.