Non-fibrogenic high mannuronate alginate coated transplants, processes for their manufacture, and methods for their use

• US 5,429,821 A
• Filed: 05/29/1992
• Issued: 07/04/1995
• Est. Priority Date: 05/29/1992
• Status: Expired due to Term

• Current Assignee: Islet Technology, Inc., Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Cochrum, Kent C., Jemtrud, Susan A., Dorian, Randel E.
• Primary Examiner(s): Page, Thurman K.
• Assistant Examiner(s): AZPURU, CARLOS A
• Application Number:
  US07/891,564
• Time in Patent Office: 1,131 Days
• Field of Search: 424/422, 424/423, 424/424, 435/1, 435/182, 435/240.241, 435/240.243, 435/240.22, 435/177, 435/178, 435/179, 435/180, 128/1 R, 264/4.1, 623/11
• US Class Current: 424/424
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0205 Chemical aspects
• CPC Class Codes: A01N 1/0231 Chemically defined matrices, e.g. alginate gels, for immobilising, holding or storing cells, tissue or organs for preservation purposes; Chemically altering or fixing cells, tissue or organs, e.g. by cross-linking, for preservation purposes
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 9/1652 Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin homeopathic globules A61K9/1623
• CPC Class Codes: A61K 9/5031 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5073 having two or more different coatings optionally including drug-containing subcoatings
• CPC Class Codes: A61K 9/5089 Processes
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/36 containing ingredients of undetermined constitution or reaction products thereof , e.g. transplant tissue, natural bone, extracellular matrix isolated natural compounds, e.g. collagen A61L27/24
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: B01J 2/003 followed by coating of the granules to prevent the granules sticking together B01J2/30
• CPC Class Codes: C08L 5/04 Alginic acid; Derivatives thereof
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 2500/12 Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
• CPC Class Codes: C12N 2533/74 Alginate
• CPC Class Codes: C12N 5/0012 Cell encapsulation
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: Y10S 623/92 Method or apparatus for preparing or treating prosthetic

Abstract

A transplant comprising a core of a viable, physiologically active cells coated with a non-fibrogenic alkaline earth metal alginate free from fibrogenic amounts of fucose, sulfate, phloroglucinol and protein moieties, having a mannuronate to guluronate molar ratio of from 1.2 to 6. The coating protects the core from host immunological destruction after transplantation. The coating is sufficiently permeable to permit the diffusion of nutrients and cell products through the coating.

A process for coating the transplant core comprising coating the core with alginate substantially free from fibrogenic compounds and reacting the alginate coating with alkaline earth metal cations comprising calcium ions or magnesium ions or mixture thereof to form an alkaline earth metal alginate coating.

Claims

1. A viable, physiologically active and acceptable tissue transplant suitable for long-term transplantation,

wherein said transplant is protected from destruction by the host immune system by coating said transplant essentially with a purified non-fibrogenic and non-toxic alkaline earth metal alginate selected from the group consisting of calcium alginate, magnesium alginate and a mixture thereof;

wherein said alkaline earth metal alginate is substantially free from fucose, sulfate, phlorogluoinol or protein moieties;

wherein said alkaline earth metal alginate provides a barrier having a thickness between about 20 μ

m and about 200 μ

m providing a prolonged protection of said transplant from destruction of said transplant by the host immune system;

wherein said barrier is permeable enough to allow diffusion and release of proteins having an intermediate size from the transplant into the host body.

8. The transplant of claim 1 wherein the concentration of fucose moieties is less than 1 wt. %, of sulfate moieties is less then 0.5 wt. %, and of phloroglucinol moieties is less than 0.01 wt. %.

9. The transplant of claim 8 wherein the alkaline earth metal alginate contains mannuronate and guluronate in a molar ratio of from 1.2 to 6.

10. The transplant of claim 9 wherein the transplant material is selected from pancreatic islet cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid cells, adrenal cells, thymic cells, ovarian cells and hepatic cells.

11. The transplant of claim 10 wherein the alkaline earth metal is calcium.

12. The transplant of claim 11 additionally comprising a polylysine-alginate complex on the outer surface thereof.

13. The transplant of claim 12 wherein the polylysine-alginate complex further comprises polyaspartic acid.

14. The transplant of claim 12 wherein the alginate coating is at least partially liquified and wherein the polylysine-alginate complex further comprises polylysine.

15. The transplant of claim 14 wherein the transplant is pancreatic islet cells.

2. A viable, physiologically active and acceptable pancreatic islet transplant suitable for long-term transplantation,

wherein said transplant is protected from destruction by the host immune system by coating said transplant essentially with a purified non-fibrogenic and non-toxic alkaline earth metal alginate selected from the group consisting of calcium alginate, magnesium alginate and a mixture thereof;

wherein said alkaline earth metal alginate is substantially free from fucose, sulfate, phloroglucinol or protein moieties;

wherein said alkaline earth metal alginate provides a barrier having a thickness between about 20 μ

m and about 200 μ

m providing a prolonged protection of said transplant from destruction by the host immune system;

wherein said barrier is permeable enough to allow diffusion and release of pancreatic hormones from the transplant into the host body.

3. The pancreatic islets transplant of claim 2 wherein the concentration of fucose moieties is less than 1 wt. %, of sulfate moieties is less then 0.5 wt. %, and of phloroglucinol moieties is less than 0.01 wt. %.

4. The pancreatic islets transplant of claim 3 wherein the alkaline earth metal alginate contains mannuronate and guluronate in a molar ratio of from 1.2 to 6.

5. The pancreatic islets transplant of claim 4 additionally comprising a polylyeine-alginate complex on the outer surface thereof.

6. The pancreatic islets transplant of claim 5 wherein the polylysine-alginate complex further comprises polyaspartic acid.

7. The pancreatic islets transplant of claim 5 wherein the alginate coating is at least partially liquified and wherein the polylysine-alginate complex further comprises polylysine.

16. A process of making a non-fibrogenic, coated core of transplant material suitable for transplantation from a donor to a host comprising:

a) coating the transplant material with an alginate which is free from fibrogenic concentrations of fucose, sulfate, phloroglucinol and protein moieties, and

b) reacting the alginate coating with alkaline earth metal cations to form an alkaline earth metal alginate coating.

17. The process of claim 16 wherein the alkaline earth metal cations are selected from the group consisting of calcium ions, magnesium ions, or mixtures thereof.

18. The process of claim 16 wherein, in step a, the concentration of fucose moieties is less than 1 wt. %, of sulfate moieties is less then 0.5 wt. %, and of phloroglucinol moieties is less than 0.01 wt. %.

19. The process of claim 16 wherein the alginate has a mannuronate to guluronate molar ratio of from 1.2 to 6.

20. The process of claim 16 wherein the alkaline earth metal cation is a calcium ion.

21. The process of claim 16 wherein the alginate coating has a permeability sufficiently low and a thickness sufficiently large to protect the tissue cells from host immunological agents after transplantation, the coating also being sufficiently permeable and thin to permit the diffusion of cell nutrients and cell products through the coating sufficient for cell viability.

22. The process of claim 21 wherein the coating has a thickness between at least about 20 μ

m and about 200 μ

m.

23. The process of claim 16 wherein the transplant is material selected from the group consisting of pancreatic islet cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid cells, adrenal cells, thymic cells, ovarian cells and hepatic cells.

24. The process of claim 16 wherein the transplant material is pancreatic islet cells.

25. The process of claim 16 further comprising

c) reacting the alkaline earth metal alginate coating with an aqueous solution containing sufficient polylysine to form a polylysine-alginate complex on the outer surface of said coatings.

26. The process of claim 25 further comprising

d) reacting the polylysine-alginate complex with an amount of polyaspartic acid sufficient to negatively charge the coating.

27. The process of claim 25 further comprising

d) reacting the polylysine-alginate complex coating with an alkaline earth metal sequestering agent to at least partially liquify the alginate, wherein liquified alginate reacts with available reaction sites of the polylysine, thereby negatively charging the coating.

28. The process of claim 16 further comprising a pretreatment step of contacting said transplant material with polylysine prior to step a.