Growth of adult pancreatic islet cells

• US 5,681,587 A
• Filed: 03/29/1996
• Issued: 10/28/1997
• Est. Priority Date: 10/06/1995
• Status: Expired due to Fees

• Current Assignee: CyThera, Inc. (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Desmos, Inc.
• Inventors: Zimber, Michael, Halberstadt, Craig, Grzesiak, John J.
• Primary Examiner(s): Chan, Christina
• Assistant Examiner(s): VANDERVEGT, FRANCOIS P
• Application Number:
  US08/626,394
• Time in Patent Office: 578 Days
• Field of Search: 424/520, 424/550, 424/556, 424/562, 424/572, 435/240.2, 435/240.21, 435/240.23, 435/350, 435/351, 435/352, 435/363, 435/366, 435/395, 435/402, 514/3, 514/4
• US Class Current: 424/562
• 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: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

A method of increasing the number of adult pancreatic islet cells available for transplantation by contacting the cells with laminin 5 extracellular matrix. When contacted with the deposited matrix produced by 804G rat bladder carcinoma cells, a 1,500 fold increase in cell number is observed after three passages in culture. Islet cell expansion also occurs when cells are contacted with 804G soluble matrix. The expanded islet cells contain insulin and respond to glucose challenge.

Claims

1. A method of expanding adult pancreatic islet-like cell clusters (ICCs), comprising the steps of:

culturing said ICCs in contact with laminin 5; and

passaging said cultured ICCs in contact with laminin 5 at least once, whereby the number of said cultured ICCs is significantly increased after said passaging.

2. The method of claim 1, wherein said laminin 5 is the extracellular matrix produced by 804G, ATCC 11555 or NBT-II, ATCC 11556, rat bladder carcinoma cells, said matrix comprising three polypeptides having molecular weights of about 150 kD, 140 kD and 135 kD, said matrix characterized as:

(a) promoting enhanced growth of said ICCs in comparison to ICCs grown in the absence of the matrix;

(b) having the ability to promote hemidesmosome formation in epithelial cells cultured thereon;

(c) binding concanavalin; and

(d) being bound by polyclonal antibodies generated against the matrix.

3. The method of claim 1, wherein said ICCs are from a mammal.

4. The method of claim 3, wherein said ICCs are from a human.

5. The method of claim 2, wherein said laminin 5 is deposited by said 804G, ATCC 11555 or said NBT-II, ATCC 11556, cells onto a substrate, said cells are removed from said matrix, and said ICCs are grown in contact with said matrix.

6. The method of claim 2, wherein said matrix is secreted by said 804G, ATCC 11555 or said NBT-II, ATCC 11556, cells into said culture media.

7. The method of claim 6, further comprising purifying said secreted laminin 5 from said culture media.

8. The method of claim 2, wherein said laminin 5 is produced by 804G, ATCC 11555, cells.

9. The method of claim 2, wherein said polypeptides are produced from recombinant DNA.

10. The method of claim 9, wherein said recombinant DNA is human.

11. The method of claim 1, wherein said laminin 5 is selected from the group consisting of kalinin and epiligrin.

12. The method of claim 1, wherein said laminin 5 is MCF 10A, ATCC CRL-10317, matrix.

13. The pancreatic ICCs prepared in accordance with claim 1.

14. The method of claim 1, wherein said ICCs are expanded at least 10 fold.

15. The method of claim 1, wherein said ICCs are expanded at least 100 fold.

16. The method of claim 1, wherein said ICCs in contact with laminin 5 are passaged at least twice, and obtaining functional insulin-producing passaged cells.

17. A method of treating Type I diabetes in a patient in need thereof, comprising the step of administering to said patient an effective insulin-producing amount of the ICCs of claim 13.

18. The method of claim 17, wherein said administering step is by implantation under the kidney capsule.

19. The method of claim 18, wherein said ICCs are placed in an immunoprotective barrier prior to said implantation.

20. The method of claim 17, wherein said administering step is by direct injection into the liver.

21. The method of claim 17, wherein said effective insulin-producing amount is between about 2×

10⁵ and about 8×

10⁵ ICCs.