Preparing artificial organs by forming polylayers of different cell populations on a substrate

• US 6,428,802 B1
• Filed: 12/29/1999
• Issued: 08/06/2002
• Est. Priority Date: 12/29/1999
• Status: Expired due to Term

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Atala, Anthony
• Primary Examiner(s): Naff, David M.
• Application Number:
  US09/474,524
• Time in Patent Office: 951 Days
• Field of Search: 435/177, 435/180, 435/395, 424/423, 424/93.7
• US Class Current: 424/423
• CPC Class Codes: A61F 2/00 Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents as cosmetic articles, see the relevant subclasses, e.g. wigs, hair pieces, A41G3/00, A41G5/00; artificial nails A45D31/00; dental prostheses A61C13/00; materials for prostheses A61L27/00; artificial hearts A61M60/00; artificial kidneys A61M1/14
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3891 as distinct cell layers
• CPC Class Codes: C12N 2502/1347 Smooth muscle cells
• CPC Class Codes: C12N 2502/253 Bladder cells
• CPC Class Codes: C12N 5/0685 Bladder epithelial cells
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

Compositions and methods are provided for producing multilayered artificial organs comprising heterogenous polylayers. Polylayers comprising homogenous cell populations are created on one side of a biocompatible substrate such that a chimeric interface is produced between the heterogenous polylayers. Cellular interaction at the chimeric interface produce an interstitial biomaterial with morphological and functional characteristics that resemble the natural in vivo organ. An artificial organ is produced by creating a first cultured polylayer of cells derived from an isolated population of smooth muscle cells on a substrate in the shape of an organ, and creating a second cultured polylayer of cells derived from a cell population different from the smooth muscle cell population.

Claims

1. An artificial organ construct comprising:

a biocompatable substrate;

a first cultured polylayer of cells derived from an isolated population of smooth muscle cells on the biocompatible substrate; and

a second cultured polylayer of cells derived from a second cell population that is different from the smooth muscle cell population, wherein the second polylayer is coupled to the first polylayer at a boundary between the first and second polylayer such that the construct provides the functional equivalent of a natural biological structure upon implantation.

2. The artificial organ of claim 1, further comprising a third cultured polylayer of cells derived from a third cell population that is different from the smooth muscle cell population and the second cell population, wherein the third polylayer is coupled to the second polylayer at a boundary between the second and third polylayer.

3. The artificial organ of claims 1 or 2, wherein the at a boundary between the polylayers further comprises an interstitial biomaterial produced by at least one of the polylayers.

4. The artificial organ of claim 3, wherein the interstitial biomaterial comprises cells with a normal morphology.

5. The artificial organ of claim 1, further comprising layering factors between the first and second polylayers, wherein the factors are selected from the group consisting of nutrients, growth factors, cytokines, extracellular matrix components, inducers of differentiation, products of secretion, immunomodulators, and biologically-active compounds which enhance or allow growth of the cellular network or nerve fibers.

6. The artificial organ of claim 1, wherein the organ is selected from the group consisting of heart, kidney, liver, pancreas, spleen, bladder, ureter and urethra.

7. The artificial organ of claim 1, wherein the artificial organ is a part of an organ selected from the group consisting of heart, kidney, liver, pancreas, spleen, bladder, ureter and urethra.

8. An artificial bladder construct comprising:

a biocompatable substrate;

a first cultured polylayer of cells derived from an isolated population of smooth muscle cells on the biocompatable substrate; and

a second cultured polylayer of cells derived from a urothelial cell population, wherein the second polylayer is coupled to the first polylayer at a boundary between the first and second polylayer such that the construct provides the functional equivalent of a natural bladder upon implantation.

9. The artificial bladder of claim 8, wherein the boundary between the first and second polylayer further comprises an interstitial submucosa produced by at least one of the polylayers.

10. A method for producing an artificial organ construct comprising:

providing a biocompatable substrate in the shape of an organ;

creating a first cultured polylayer of cells derived from an isolated population of smooth muscle cells on the biocompatable substrate, wherein the first polylayer is attached to the biocompatable substrate; and

creating a second cultured polylayer of cells derived from a second cell population that is different from the smooth muscle cell population, wherein the second polylayer is coupled to the first polylayer at a boundary between the first and second polylayer such that the construct provides the functional equivalent of a natural biological structure upon implantation, thereby producing an artificial organ construct.

11. The method of claim 10, further comprising creating a third cultured polylayer of cells derived from a third cell population that is different from the smooth muscle cell population and the second cell population, wherein the third polylayer is coupled to the second polylayer at a boundary between the second and third polylayer.

12. The method of claims 10 or 11, wherein the boundary between the polylayers further comprises an interstitial biomaterial produced by at least one of the polylayers.

13. The method of claim 12, wherein the interstitial biomaterial comprises cells with a normal morphology.

14. The method of claim 10 further comprising layering factors between the first and second polylayers, wherein the factors are selected from the group consisting of nutrients, growth factors, cytokines, extracellular matrix components, inducers of differentiation, products of secretion, immunomodulators, and biologically-active compounds which enhance or allow growth of the cellular network or nerve fibers.

15. The method of claim 10, wherein the artificial organ is selected from the group consisting of heart, kidney, liver, pancreas, spleen, bladder, ureter and urethra.

16. The method of claim 10, wherein the artificial organ is a part of an organ selected from the group consisting of heart, kidney, liver, pancreas, spleen, bladder, ureter and urethra.

17. The method of claim 10, wherein the biocompatible substrate is a polymer.

18. The method of claim 10, wherein the biocompatable substrate is a decellularized organ produced by removing cellular content from the organ leaving a three-dimensional scaffold of connective tissue.

19. The method of claim 18, wherein the decellularized organ is selected from the group consisting of heart, kidney, liver, pancreas, spleen, bladder, ureter and urethra.

20. The method of claim 18, wherein the decellularized organ is a part of a decellularized organ selected from the group consisting of heart, kidney, liver, pancreas, spleen, bladder, ureter and urethra.

21. A method for producing an artificial bladder construct comprising:

providing a biocompatable substrate in the shape of a bladder;

creating a first cultured polylayer derived from an isolated population of smooth muscle cells on the biocompatable substrate, wherein the first polylayer is attached to the biocompatable substrate; and

creating a second cultured polylayer comprising a urothelial cell population, wherein the second polylayer is coupled to the first polylayer at a boundary between the first and second polylayer, such that the construct provides the functional equivalent of a natural bladder upon implantation, thereby producing an artificial bladder construct.

22. The method of claim 21, wherein the boundary between the first and second polylayer further comprises an interstitial submucosa produced by at least one of the polylayers.

23. The method of claim 21, wherein the biocompatible substrate is a polymer.

24. The method of claim 21, wherein the biocompatable substrate is a decellularized bladder produced by removing cellular content from a mammalian bladder leaving a three-dimensional scaffold of connective tissue.

25. The method of claim 21, wherein the biocompatible substrate is a part of a decellularized bladder.

26. A method for treating a subject with a genitourinary disorder comprising:

providing an artificial organ construct having a first cultured polylayer of cells derived from an isolated population of smooth muscle cells on a biocompatable substrate, and second cultured polylayer of a urothelial cell population, wherein the second polylayer is coupled to the first polylayer at a boundary between the first and second polylayer;

implanting the organ construct into the subject, such that the construct provides the functional equivalent of a natural structure; and

monitoring the subject for a modulation in the genitourinary disorder.

27. The method of claim 26, wherein the boundary between the first and second polylayer further comprises an interstitial submucosa produced by at least one of the polylayers.

28. The method of claim 26, wherein the biocompatable substrate is a polymer.

29. The method of claim 26, wherein the biocompatable substrate is a decellularized organ produced by removing cellular content from the organ leaving a three-dimensional scaffold of connective tissue.

30. The method of claim 29, wherein the decellularized organ is selected from the group consisting of bladder, ureter and urethra.

31. The method of claim 29, wherein the decellularized organ is a part of a decellularized organ selected from the group consisting of bladder, ureter and urethra.