USE OF BONE MARROW CELLS FOR LONG TERM CULTURE OF PANCREATIC ISLET CELLS

• US 20100143306A1
• Filed: 11/20/2007
• Published: 06/10/2010
• Est. Priority Date: 11/21/2006
• Status: Abandoned Application

• Current Assignee: Prospect Chartercare LLC (The Centurion Foundation, Inc.)
• Original Assignee: Roger Williams Hospital (Boston University)
• Inventors: Luo, Luguang
• Application Number:
  US12/515,754
• Publication Number:
  US 20100143306A1
• Time in Patent Office: Days
• US Class Current: 424/93.300
• 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: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2502/1394 Bone marrow stromal cells; whole marrow
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

Bone marrow cells have been demonstrated to be useful in the maintenance of pancreatic islet β cell viability, structure, and/or function in culture for a sustained period. Bone marrow cells were also found to promote β cell growth while reducing inflammatory cytokine release and reduce apoptosis. Moreover, islet cells co-cultured with bone marrow cells were shown to retain insulin response function and to function in an islet cell transplant in a mouse model of diabetes to restore normal insulin secretion. Cord blood cells and isolated peripheral CD34+ blood cells were unable to support β islet cell growth or increase survival.

Claims

1. A method for culturing islet β

cell comprising culturing an islet β

cell with a bone marrow cell in vitro.

2. The method of claim 1, wherein the islet β

cell is cultured for a sustained period.

3. The method of claim 2, wherein the sustained period is at least about 30 days, at least about 60 days, at least about 90 days or at least about 120 days.

7. The method of claim 1, wherein culturing an islet β

cell with a bone marrow cell increases islet β

cell viability, improves islet β

cell function, and/or improves islet β

cell morphology as compared to an islet β

cell cultured in the absence of a bone marrow cell.

8. The method of claim 7, wherein cell function is determined by a basal insulin secretion assay or a glucose induced insulin secretion assay.

10. The method of claim 7, wherein increased islet β

cell viability, improved islet β

cell function, and/or improved islet β

cell morphology are achieved for a sustained period.

11. The method of claim 1, wherein the method reduces apoptosis of an islet β

cell in culture with a bone marrow cell as compared to an islet β

cell cultured in the absence of a bone marrow cell.

12. The method of claim 1, wherein the method reduces cytokine release from an islet β

cell in culture with a bone marrow cell as compared to an islet β

cell cultured in the absence of a bone marrow cell.

13. The method of claim 12, wherein the cytokine is IL-1 β

.

14. The method of claim 1, wherein the method increases the expression of at least one endocrine cell specific genes.

15. The method of claim 14, wherein the endocrine cell specific genes are selected from the group consisting of GCG (glucagon, α

-cell gene), INS (insulin, β

-cell gene), and SST (somatostatin, γ

-cell gene);

transcription factors for β

cell pancreatic and duodenal homeobox1 (PDX1 or IPF1), Neurogenin 3 (NGN3), paired box gene 6 (PAX6), islet-1 (ISL1), v-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MAFa), and Mist 1.

42. A kit comprising a bone marrow cell and instructions for use in accordance with the method of claim 1.

4-6. -6. (canceled)

9. (canceled)

16. A method for pancreatic islet β

cell expansion in vitro comprising, culturing an islet β

cell with a bone marrow cell.

17. A method for increasing islet β

cell viability, improved islet β

cell function, and/or improved β

cell morphology comprising culturing an islet β

cell with a bone marrow cell in vitro.

18-24. -24. (canceled)

25. A method of treating a subject in need of an islet β

cell transplant comprising culturing an islet β

cell with a bone marrow cell in vitro; and

implanting the islet cell and the bone marrow cell into the individual in need of an islet β

cell transplant.

26-30. -30. (canceled)

31. A method of treating diabetes in a subject in need thereof comprising:

culturing an islet β

cell with a bone marrow cell in vitro; and

implanting the cultured islet β

cell into the subject;

thereby treating the subject for diabetes.

32-36. -36. (canceled)

37. A method for implanting islet β

cells in a subject comprising;

culturing an islet β

cell with a bone marrow cell in vitro; and

implanting the cultured islet β

cell into the subject.

38-41. -41. (canceled)