Preparation and xenotransplantation of porcine islets

• US 7,122,177 B2
• Filed: 01/19/2001
• Issued: 10/17/2006
• Est. Priority Date: 01/20/2000
• Status: Expired due to Term

• Current Assignee: Diatranz Otsuka Limited (Otsuka Holdings Company Limited)
• Original Assignee: Diabcell Pty Limited
• Inventors: Calafiore, Riccardo, Elliott, Robert Bartlett, Basta, Gusseppe
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US09/857,325
• Publication Number:
  US 20030044391A1
• Time in Patent Office: 2,097 Days
• Field of Search: 435/325, 435/375, 435/376, 435/378, 435/379, 435/381, 424/93.1, 424/93.7
• US Class Current: 424/93.1
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• 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: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

The invention relates to developments in the treatment of diabetes in mammals. Particularly it relates to a method of preparing a xenotransplantable porcine islet preparation capable upon xenotransplantation of producing porcine insulin in an appropriate recipient mammal, the method including or comprising the steps of:

• • (I) harvesting the pancreas of piglets at or near full term gestation, and
  • (ii) extracting pancreatic β islet cells from the harvested pancreas wherein the islets (at least at some stage in the performance of the method) are exposed to nicotinamide.

Further, the invention relates to a method of encapsulation of a xenotransplantable porcine islet preparation, and transplantation of such a preparation, or a capsule containing such a preparation, into an appropriate recipient mammal.

Claims

1. A method of preparing a xenotransplantable islet comprising in sequential order the steps of:

(i) harvesting the pancreas of a piglet, said piglet having an age of between −

20 to +10 days relative to full-term gestation,

(ii) exposing said harvested pancreas to nicotinamide,

(iii) exposing said harvested pancreas to lignocaine; and

(iv) extracting pancreatic β

islet cells from said harvested pancreas and simultaneously contacting said pancreatic β

islet cells with a quinolone antibiotic, wherein said method results in said xenotransplantable islet.

9. The method of any one of claims 1–

4, wherein the piglet has an age of between −

7 and +10 days relative to full term gestation.

10. The method of any one of claims 1–

4, wherein the step of extraction includes the use of a suitable human collagenase.

11. The method of any one of claims 1–

4, wherein the harvested pancreas is bathed in a mammalian albumin solution substantially free of microbiological agents.

12. The method of any one of claim 11, wherein the mammalian albumin comprises human serum albumin (HSA).

13. The method of any one of claims 1–

4, further comprising a step of contacting the β

islet cells with a compound selected from the group consisting of insulin-like growth factor 1 (IGF-1) and the N-terminal tripeptide of IGF-1.

14. The method of claim 13, wherein the compound consists of the N-terminal tripeptide of IGF-1.

2. A method of preparing a xenotransplantable porcine islet, said method comprising in sequential order at least the steps of:

(i) harvesting a pancreas of a piglet, the piglet having an age of between −

20 to +10 days relative to full-term gestation,

(ii) exposing said harvested pancreas to nicotinamide, and

(iii) extracting pancreatic β

islet cells from said harvested pancreas and simultaneously contacting said pancreatic β

islet cells with ciprofloxacin;

wherein said method results in said xenotransplantable porcine islet.

3. A method of preparing a xenotransplantable porcine islet, said method comprising in sequential order at least the steps of:

(i) harvesting a pancreas of a piglet, said piglet having an age of between −

20 to +10 days relative to full-term gestation,

(ii) exposing said harvested pancreas to nicotinamide, and

(iii) extracting pancreatic β

islet cells from said harvested pancreas and simultaneously contacting said pancreatic β

islet cells with lignocaine, wherein said method results in said xenotransplantable porcine islet.

4. A method of preparing a xenotransplantable porcine islet comprising in sequential order the steps of:

(i) harvesting the pancreas of a piglet, the piglet having an age of between −

20 to +10 days relative to full-term gestation,

(ii) exposing said harvested pancreas to nicotinamide,

(iii) exposing said harvested pancreas to ciprofloxacin; and

(iv) extracting pancreatic β

islet cells from said harvested pancreas and simultaneously contacting said pancreatic β

islet cells with a trauma-protecting agent, wherein said method results in said xenotransplantable islet.

15. The method of claim 4, wherein the trauma protecting agent comprises an anesthetic agent.

16. The method of claim 15, wherein the anesthetic agent comprises a phospholipase A2 inhibitor.

17. The method of claim 16, wherein the phospholipase A2 inhibitor comprises lignocaine.

5. A method of preparing a xenotransplantable porcine islet, comprising in sequential order the steps of:

(i) harvesting the pancreas of a piglet, the piglet having an age of between −

20 to +10 days relative to full term gestation,

(ii) exposing the harvested pancreas to a trauma-protecting agent lignocaine,

(iii) exposing the harvested pancreas to nicotinamide, and

(iv) extracting pancreatic β

islet cells from the harvested pancreas and simultaneously contacting the pancreatic β

islet cells with a quinolone antibiotic;

the method resulting in a xenotransplantable islet.

6. A method of preparing a xenotransplantable porcine islet comprising in sequential order the steps of:

(i) harvesting the pancreas of a piglet, the piglet having an age of between −

20 to +10 days relative to full term gestation,

(ii) exposing the harvested pancreas to the quinolone antibiotic ciprofloxacin,

(iii) exposing the harvested pancreas to nicotinamide, and

(iv) extracting pancreatic β

islet cells from the harvested pancreas and simultaneously contacting the pancreatic β

islet cells with a trauma protecting agent lignocaine;

the method resulting in a xenotransplantable islet.

7. A method of preparing a xenotransplantable porcine islet comprising in sequential order the steps of:

(i) harvesting the pancreas of a piglet, said piglet having an age of between −

20 to +10 days relative to full-term gestation;

(ii) exposing said harvested pancreas of said piglet to nicotinamide;

(iii) exposing said harvested pancreas to lignocaine;

(iv) extracting pancreatic β

islet cells from said harvested pancreas in the presence of a suitable collagenase; and

(v) contacting said extracted pancreatic β

islet cells with a quinolone antibiotic, wherein said method results in the preparation of said xenotransplantable porcine islet.

8. A method of preparing a xenotransplantable porcine islet comprising in sequential order the steps of:

(i) harvesting the pancreas of a piglet, said piglet having an age of between −

20 to +10 days relative to full-term gestation;

(ii) exposing said harvested pancreas of said piglet to nicotinamide;

(iii) exposing said harvested pancreas to a trauma-protecting agent;

(iv) extracting pancreatic β

islet cells from said harvested pancreas in the presence of a suitable collagenase; and

(v) contacting said extracted pancreatic β

islet cells with the quinolone antibiotic ciprofloxacin, wherein said method results in the preparation of said xenotransplantable porcine islet.