Coated transplant and method for making same

• US 5,470,731 A
• Filed: 06/08/1994
• Issued: 11/28/1995
• Est. Priority Date: 05/29/1992
• Status: Expired due to Term

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Cochrum, Kent C.
• Primary Examiner(s): Azpuru, Carlos A.
• Application Number:
  US08/255,577
• Time in Patent Office: 538 Days
• Field of Search: 424/423, 424/424, 435/1, 435/177, 435/182, 435/240, 435/241, 435/240.22, 435/240.243, 264/4, 427/2.1
• US Class Current: 435/182
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 9/1641 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
• 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: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/32 Polylysine, polyornithine
• CPC Class Codes: C12N 2533/76 Agarose, agar-agar
• CPC Class Codes: C12N 5/0012 Cell encapsulation
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

Transplants are coated with an insoluble immunological barrier membrane. The membrane can comprise a non-cytotoxic first layer of agarose and cationic amino acid polymer and a non-cytotoxic second layer chemically bonded to said first layer, the second layer comprising an anionic amino acid polymer. The molecular weight of the anionic amino acid polymer is preferably in the range of 2000 to 500,000 daltons, and the thickness if the total membrane coating is within the range of 1 and 100 microns. Alternatively, the membrane can comprise a non-cytotoxic first layer of a cationic amino acid polymer. A non-cytotoxic second layer chemically bonded to the first layer comprises agarose and an anionic amino acid polymer. A non-cytotoxic optional third layer chemically bonded to said second layer comprises a cationic amino acid polymer. The molecular weight of the cationic acid polymer of the third layer is in the range of from 2,000 to 500,000 daltons and the thickness of the total coating is between 1 and 100 microns. The transplants can be allografts of endocrine cells or tissues such a pancreatic islets. The preferred cationic amino acids are L-lysine, poly-L-arginine, and mixtures thereof, and the preferred anionic amino acids are L-aspartic acid, L-glutamic acid, and mixtures thereof. The coatings are formed by applying corresponding successive solutions to the cells or tissue of the transplant and by forming droplets in immiscible oils.

Claims

1. A method for coating a tissue transplant with an immunological barrier protecting the transplant against destruction by the immune system of an immunologically incompatible transplant host recipient, said method comprising steps:

(a) isolating the transplant tissue;

(b) coating the transplant tissue with a first coating by suspending the transplant in an aqueous solution of a mixture of agarose and a cationic or anionic non-cytotoxic amino acid polymer at a temperature which renders the agarose to be in a liquid sol state;

(c) lowering a temperature of the suspension of step (b) until the agarose changes from the liquid sol to a gel;

(d) coating the transplant with a second coating by dispersing the suspension of step (c) in a solution of an cationic or anionic amino acid polymer having the charge opposite to the amino acid polymer used in the step (b), and chemical bonding the amino acid of the first coating with the amino acid of the second coating, said chemical bonding formed by one or more covalent, ionic or hydrogen bonds between the first and second layer.

2. The method of claim 1 wherein the first coating contains agarose from about 0.5% to about 8%, by weight, and the cationic or anionic amino acid from about 0.1% to 1%, by weight.

3. The method of claim 2 wherein the first coating contains agarose from about 0.8% to about 1.4%, by weight.

4. The method of claim 3 wherein the temperature of step (b) rendering the agarose to be in a liquid sol state is above the agarose gelling temperature and up to about 37°

C.

5. The method of claim 4 wherein the first coating mixture of step (b) is chilled to an agarose gelling temperature from about 5°

C. to about 35°

C. changing the agarose liquid sol into the agarose gel.

6. The method of claim 6 wherein the mixture of step (b) containing the transplant coated with agarose sol is chilled by introducing said mixture into a noncytotoxic, pharmaceutically acceptable oil at a temperature above the agarose gelling temperature and chilling said mixture with continuous stirring to the gelling temperature of the agarose.

7. The method of claim 6 wherein the dispersion of step (d) contains from about 0.1% to about 1%, by weight, of the amino acid polymer.

8. The method of claim 7 wherein the cationic amino acid is selected from the group consisting of L-lysine, L-arginine and a mixture thereof, and the anionic amino acid is selected from group consisting of L-aspartic acid, L-glutamic acid and a mixture thereof.

9. The method of claim 8 wherein the tissue transplant is an allograft.

10. The method of claim 8 wherein the tissue transplant is an endocrine tissue transplant.

11. The method of claim 8 wherein the tissue transplant is a pancreatic islet, wherein the amino acid polymer present in the aqueous agarose solution used for the first coating is poly-L-lysine and wherein the amino acid polymer used for the second coating is poly-L-aspartic acid.

12. The method of claim 8 wherein the cationic amino acid polymer is poly-L-lysine.

13. The method of claim 8 wherein the anionic amino acid polymer is poly-L-aspartic acid.

14. The method of claim 11 wherein the pancreatic islet transplant is additionally coated with a third layer consisting of the poly-L-lysine.

15. A method for coating a tissue transplant with an immunological barrier protecting the transplant against destruction by the immune system of an immunologically incompatible transplant host recipient, said method comprising steps:

(a) isolating the transplant tissue;

(b) coating the transplant tissue with a first coating comprising a solution consisting of about 0.1% to about 1%, by weight, of a cationic or anionic acid polymer, said amino acid polymer having a charge opposite to a charge on a transplant surface, reacting with the transplant surface charge and forming a chemical bond;

(c) dispersing the transplant of the step (b) in a second coating solution comprising a mixture of agarose and a cationic or anionic amino acid polymer having charge opposite to the charge of the amino acid polymer of step (b), at a temperature which renders the agarose to be in a liquid sol state; and

(d) lowering the temperature of the aqueous solution of step (b) until the agarose changes from the liquid sol to a gel.

16. The method of claim 15 wherein the second coating contains agarose from about 0.5% about 8% by weight and the cationic or anionic amino acid is present in 0.1% to 1% by weight.

17. The method of claim 16 wherein the first coating contains agarose about 0.8% about 1.4%.

18. The method of claim 17 wherein the temperature of step (c) rendering the agarose to be in a liquid sol state is above the agarose gelling temperature and up to about 37°

C.

19. The method of claim 18 wherein the second coating mixture of step (c) is chilled to an agarose gelling temperature changing the agarose liquid sol into the agarose gel.

20. The method of claim 19 wherein the tissue transplant is an allograft.

21. The method of claim 19 wherein the tissue transplant is an endocrine tissue transplant.

22. The method of claim 19 wherein the tissue transplant is a pancreatic islet, wherein the amino acid polymer used for the first coating is poly-L-lysine and wherein the amino acid polymer present in the aqueous agarose solution used for the second coating is poly-L-aspartic acid.