Growing xenotransplant material in culture

• US 20050042746A1
• Filed: 09/27/2002
• Published: 02/24/2005
• Est. Priority Date: 09/28/2001
• Status: Abandoned Application

• Current Assignee: Diabcell Pty Limited
• Original Assignee: Diabcell Pty Limited
• Inventors: Garkavenko, Olga
• Application Number:
  US10/490,717
• Publication Number:
  US 20050042746A1
• Time in Patent Office: Days
• US Class Current: 435/325
• 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: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 2502/22 pancreatic cells
• CPC Class Codes: C12N 2502/246 Cells of the male genital tract, non-germinal testis cells
• CPC Class Codes: C12N 5/0677 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: C12N 5/0683 Cells of the male genital tract, e.g. prostate, epididymis; Non-germinal cells from testis, e.g. Leydig cells, Sertoli cells spermatogonia C12N5/061
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

Over time, cultured mammalian cells aggregate into an artificial tissue with 10% homologous serum and nicotinamide. Neonatal porcine islets alone have a SGS index of 4.6. Co-culture with neonatal porcine Sertoli cells formed free-floating islet-like structures, 300-600 microns in diameter (SGS index 21). Basal secretion of insulin remains high for islets in a heavily confluent mixed cell culture. Re-seeded cultures were very responsive to glucose (SGS index 21.8). Co-culture with fibroblasts also provides a high SGS index, permitting use of the recipient'"'"'s fibroblast line as support/trophic cells for an xenotransplantable graft, as for treatment of diabetes.

Claims

1. A method for carrying out long-term in vitro assembly, propagation and maturation of a functional artificial tissue comprised of more than one kind of cell taken from at least one species of mammal, characterised in that a selected first type of cell capable of being stimulated to secrete a product is co-cultured over an extended period together with at least one selected second type of cell capable of providing a support/trophic function for the first type, in a growth medium supplemented with an effective amount of homologous serum from the species of mammal that provided the first type of cell, so that organised functional, synergistic groups of mixed cell types are developed over the extended period.

2. A method as claimed in to claim 1 for making a functional artificial tissue characterised in that the artificial tissue includes a first type of cell and a second type of cell jointly capable of possessing a functional property on being co-cultured in a growth media supplemented with an effective amount of homologous serum, so that the artificial tissue may be grafted into a recipient suffering from a deficiency of the property, to alleviate the deficiency.

3. A method as claimed in claim 1, characterised in that the first type of cell is selected from the group of endocrine secretory cells and the product is an endocrine hormone.

4. A method as claimed in claim 3, characterised in that the second type of cell is selected from a group including Sertoli cells and fibroblasts.

5. A method as claimed in claim 1, characterised in that the effective amount of homologous serum is in a range of from about 5% to about 15% by volume in the growth medium.

6. A method as claimed in claim 1, characterised in that the growth medium also includes nicotinamide within a range of from about 5 mM to about 15 mM.

7. A method as claimed in claim 1, characterised in that at least the first type of cell is collected from a neonatal mammal.

8. A method as claimed in claim 4 for making a functional artificial tissue characterised in that the first type of cell is endocrine cells from the pancreatic islets of Langerhans and that a ratio of the first type of cell to the second type of cell is optimised.

9. A method as claimed in claim 8 for making an artificial tissue characterised in that the artificial tissue is grown as a co-culture in a growth media supplemented with an effective amount of homologous serum, glucose, and nicotinamide, so that the resulting artificial tissue exhibits a functional secretory response to a local concentration of glucose, so that the artificial tissue may be grafted into a recipient suffering from diabetes in order to alleviate the disease.

10. A method as claimed in claim 9 further characterised in that the second type of cell is taken from an individual in need of a functional graft and after in vitro propagation and maturation together with the first type of cell for a period is returned to the diabetic recipient in the form of a functional artificial tissue.

11. A method as claimed in claim 10 further characterised in that the functional cell combinations have been grown as a co-culture for from about 30 to about 50 days in growth media of this type.

12. A functional artificial tissue comprised of more than one kind of cell taken from at least one species of mammal, characterised in that the functional artificial tissue is comprised of at least partially organised synergistic accumulations of cells comprised of a first type of cell capable of being stimulated to secrete a product, and at least one second type of cell capable of providing a support/trophic function for the first type of cell, the functional artificial tissue being formed by co-culture in vitro over an extended period of time.

13. A functional artificial tissue as claimed in claim 12, characterised in that the first type of cell is selected from the range of endocrine secretory cells, and is capable of expressing effective amounts of at least one secretion into the growth medium when in association with the second type of cell, so that the one or more secretions may be isolated from time to time for use.

14. A secreted product from a functional artificial tissue as claimed in claim 13, characterised in that the secretion is insulin.

15. A functional artificial tissue as claimed in claim 12, characterised in that the functional artificial tissue is preserved in a state of metabolic arrest suitable for transport storage and later use.