Bioartificial devices and cellular matrices therefor

• US 5,830,492 A
• Filed: 12/07/1995
• Issued: 11/03/1998
• Est. Priority Date: 02/24/1992
• Status: Expired due to Term

• Current Assignee: Encelle, Inc. (Surgalign Holdings, Inc.)
• Original Assignee: Encelle, Inc. (Surgalign Holdings, Inc.)
• Inventors: Usala, Anton-Lewis
• Primary Examiner(s): AZPURU, CARLOS A
• Application Number:
  US08/568,694
• Time in Patent Office: 1,062 Days
• Field of Search: 424/423, 424/424, 435/1.1, 435/182, 623/11, 514/772.3
• US Class Current: 424/424
• CPC Class Codes: A01N 1/02 Preservation of living parts
• 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: A61F 2/022 Artificial gland structures using bioreactors hemodialysis A61M1/16; hemofiltration A61M1/34; other extracorporeal blood treatment A61M1/36
• CPC Class Codes: A61F 2310/00365 Proteins; Polypeptides; Degradation products thereof
• CPC Class Codes: A61K 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• 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: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 9/0024 Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L
• CPC Class Codes: A61L 27/222 Gelatin
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: C12N 2500/05 Inorganic components
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 2533/70 : Polysaccharides
• CPC Class Codes: C12N 5/0012 : Cell encapsulation
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0677 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

A device for the effective release of cellular moieties, including hormones, wherein a matrix containing a hormone producing cellular moiety is encapsulated with a non-immunogenic polymeric material of poly-para-xylylene or other aromatic based moiety having a membrane portion with a porosity blocking passage therethrough of immunogenic agents and permitting passage therethrough of effective nutrients for said cellular moiety and the hormone produced thereby, an improved matrix for the storage, manufacture, functional testing, and viral infection testing of cellular moieties wherein a collagen based hydrogel is processed to present a liquid phase at host temperature and functions as a substrate for cellular attachment with additives effective for limiting thermal and pressure trauma, and an improved method for the harvesting tissue from organs.

Claims

1. A method for harvesting islets from pancreata, comprising the steps of:

digesting the pancreata with an enzymic solution until a substantial amount of islet fragments are present with whole islets in said solution;

purifying said solution to obtain a pellet of said islets and said islet fragments;

placing said pellet in a matrix comprising a gelatin in a concentration of about 0.01 to 30 mM and a polymeric buffer substrate in a concentration of 0.01 to 1000 micromolar; and

storing said matrix until said islet fragments are substantially discernible.

9. The invention as recited in claim 1 wherein said enzymic solution is a protease.

10. The invention as recited in claim 9 wherein said protease is selected from the group comprising collagenase, trypsin, Liberase, and proteolytic enzymes.

11. The invention as recited in claim 1 wherein said enzymic solution includes from about 0.001 micromolar to 300 mM of a nitric oxide inhibitor.

12. The invention as recited in claim 11 wherein said nitric oxide inhibitor is selected from the group comprising dextran, heparian, cysteine or cystine.

13. The invention as recited in claim 11 wherein said nitric oxide inhibitor is an L-arginine analogue selected from the group aminoguanidine, N-monomethyl L-arginine, N-nitro-L-arginine, or D-arginine.

2. In the digestion of tissue using an enzymic solution for releasing a desirable cellular moiety from the tissue, the improvement comprising:

adding to said enzymic solution an effective amount of a nitric oxide inhibitor.

3. The invention as recited in claim 2 wherein said effective concentration of said nitric oxide inhibitor is about 0.001 micromolar to 300 mM.

4. The invention as recited in claim 3 wherein said nitric oxide inhibitor is a sulfated moiety.

5. The invention as recited in claim 4 wherein said sulfated moiety is selected from the group comprising dextran, heparin, cysteine or cystine.

6. The invention as recited in claim 2 wherein said nitric oxide inhibitor is a nitric oxide synthase.

7. The invention as recited in claim 3 wherein said nitric oxide inhibitor is an L-arginine analogue.

8. The invention as recited in claim 7 wherein said L-arginine analogue is selected from the group comprising aminoguanidine, N-monomethyl L-arginine, N-nitro-L-arginine, or D-arginine.

14. A method for long term storage of harvested islets from pancreata, said method comprising placing said harvested islets in a matrix comprising a gelatin and a polymeric buffer substrate to form a coating having a porosity to permit passage of nutrients and biological agents and prevent passage of immunogenic agents.

15. The invention as recited in claim 14, wherein said matrix further comprises a nitric oxide inhibitor.

16. The invention as recited in claim 15, wherein said nitric oxide inhibitor is selected from the group comprising dextran, heparian, cysteine or cystine.

17. The invention as recited in claim 15, wherein said nitric oxide inhibitor is an L-arginine analogue selected from the group aminoguanidine, N-monomethyl, L-arginine, N-nitro-L-arginine, or D-arginine.

18. The invention as recited in claim 14, wherein said matrix further comprises a cyropreservant.

19. The invention as recited in claim 18, wherein said cryopreservant is sulfate dextran.

20. The invention as recited in claim 15, wherein said matrix further comprises a cryopreservant.

21. The invention as recited in claim 20, wherein said cryopreservant is sulfated dextran.