Method for differentiating islet precursor cells into beta cells

• US 20030109036A1
• Filed: 09/19/2002
• Published: 06/12/2003
• Est. Priority Date: 12/06/2001
• Status: Abandoned Application

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Wu, Frederick D.
• Application Number:
  US10/251,004
• Publication Number:
  US 20030109036A1
• Time in Patent Office: Days
• US Class Current: 435/366
• 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 5/0676 Pancreatic cells

Abstract

The invention includes a method of differentiating pancreatic islet stem cells or islet precursor cells into functioning beta cells to treat diabetes mellitus by transplanting the cells into a diabetic animal. Pancreatic cells are isolated and cultured such that the population of nestin-positive cells increases. The cells are then cultured on poly-D-lysine such that cell aggregates form. The cell aggregates are transplanted into a diabetic animal, where they produce insulin and lower blood glucose concentrations.

Claims

1. A method for isolating pancreatic cells capable of differentiating into beta cells, comprising:

isolating pancreatic cells;

culturing said pancreatic cells, wherein said culturing results in an increase in the number of cells that express nestin, as shown by nestin-positive staining;

culturing said nestin-positive cells on poly-D-lysine;

identifying an islet-like cell aggregate in said culture of nestin-positive cells; and

,

isolating said islet-like cell aggregate, wherein said islet-like cell aggregate will function to lower the blood glucose concentration of a diabetic animal, after transplantation into said animal.

2. The method of claim 1, wherein said pancreatic cells are isolated from a human.

3. The method of claim 1, wherein, before culturing on poly-D-lysine, said nestin-positive cells are also vimentin-positive.

4. The method of claim 1, wherein, before culturing on poly-D-lysine, said nestin-positive cells are also cytokeratin-negative.

5. The method of claim 1, wherein, before culturing on poly-D-lysine, said nestin-positive cells are also insulin-negative and glucagon-negative.

6. The method of claim 1, wherein said culturing on poly-D-lysine is done for a period of about one to two days.

7. The method of claim 1, wherein, after culturing on poly-D-lysine, said islet-like cell aggregate expresses insulin.

8. The method of claim 1, wherein the diabetic animal into which the islet-like cell aggregate is transplanted is a human.

9. The method of claim 1, wherein the diabetic animal into which the islet-like cell aggregate is transplanted is an animal whose blood glucose concentration is well-controlled.

10. A method for differentiating islet precursor cells into beta cells, comprising:

isolating pancreatic cells;

culturing said pancreatic cells to enrich the culture for nestin-positive cells;

culturing said pancreatic cells on poly-D-lysine;

identifying an islet-like cell aggregate in said culture of pancreatic cells;

isolating said islet-like cell aggregate, wherein said islet-like cell aggregate expresses insulin.

11. The method of claim 10, wherein said pancreatic cells are isolated from a human.

12. The method of claim 10, wherein said enriching of the culture for nestin-positive cells is accomplished by conscientious neglect.

13. The method of claim 10, wherein, before culturing on poly-D-lysine, said nestin-positive cells are also vimentin-positive.

14. The method of claim 10, wherein, before culturing on poly-D-lysine, said nestin-positive cells are also insulin-negative and glucagon-negative.

15. The method of claim 10, wherein a cell of said islet-like cell aggregate will function as a beta cell after transplantation into a diabetic animal.

16. The method of claim 15, wherein the diabetic animal into which the islet-like cell aggregate is transplanted is a human.

17. The method of claim 15, wherein the diabetic animal into which the islet-like cell aggregate is transplanted is an animal whose blood glucose concentration is well-controlled.