Identification and high-yield isolation of human pancreatic islet progenitor and stem cells

• US 20040005301A1
• Filed: 02/10/2003
• Published: 01/08/2004
• Est. Priority Date: 02/12/2002
• Status: Abandoned Application

• Current Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Original Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Keyoung, Hansoo Michael, Goldman, Steven A.
• Application Number:
  US10/364,029
• Publication Number:
  US 20040005301A1
• Time in Patent Office: Days
• US Class Current: 424/93.21
• 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 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells

Abstract

The present invention relates to a method of separating pancreatic Islet cells or progenitor or stem cells thereof from a mixed population of cells from the pancreas. This involves selecting an enhancer/promoter which functions in the pancreatic Islet cells or progenitor cells thereof and introducing a nucleic acid molecule encoding a fluorescent protein under control of the enhancer/promoter into the mixed population of cells. The pancreatic Islet cells or progenitor or stem cells thereof are then allowed to express the fluorescent protein. The fluorescent cells (i.e. the said pancreatic Islet cells or progenitor or stem cells thereof) are then separated from the mixed population of cells. Also disclosed is an enriched or purified preparation of isolated human pancreatic Islet cells or progenitor or stem cells and the use of these cells in a method of treating a diabetic condition by transplanting the cells into a subject.

Claims

1. A method of separating pancreatic Islet cells or progenitor or stem cells thereof from a mixed population of cells from the pancreas comprising:

selecting an enhancer/promoter which functions in said pancreatic Islet cells or progenitor cells thereof;

introducing a nucleic acid molecule encoding a fluorescent protein under control of said enhancer/promoter into the mixed population of cells;

allowing the pancreatic Islet cells or progenitor or stem cells thereof to express the fluorescent protein; and

separating the fluorescent cells from the mixed population of cells, wherein said separated cells are said pancreatic Islet cells or progenitor or stem cells thereof.

2. The method of claim 1, wherein said introducing comprises viral mediated transduction of said pancreatic Islet stem cells and progenitors thereof.

3. The method of claim 2, wherein said viral mediated transduction comprises adenovirus-mediated transduction.

4. The method of claim 2, wherein said viral mediated transduction comprises retroviral-mediated transduction.

5. The method of claim 2, wherein said viral mediated transduction comprises lentiviral-mediated transduction.

6. The method of claim 2, wherein said viral mediated transduction comprises transduction by adeno-associated virus.

7. The method of claim 1, wherein said introducing comprises electroporation.

8. The method of claim 1, wherein said introducing comprises naked DNA insertion.

9. The method of claim 8, wherein said introducing comprises biolistic penetration.

10. The method of claim 1, wherein said introducing comprises liposomal-mediated transformation of said mixed population of cells.

11. The method of claim 1, wherein said separating comprises fluorescence activated cell sorting.

12. The method of claim 1, wherein said enhancer/promoter is an E/nestin enhancer/promoter.

13. The method of claim 1, wherein said enhancer/promoter is a Musashi promoter.

14. The method of claim 1, wherein said enhancer/promoter is an NKX6.1 promoter.

15. The method of claim 1, wherein said enhancer/promoter is a neurogenin-3 promoter.

16. The method of claim 1, wherein said enhancer/promoter is an HB9 promoter.

17. The method of claim 1, wherein said enhancer/promoter is an PDX-1 promoter.

18. The method of claim 1, wherein the mixed population of cells is derived from pancreatic tissue.

19. The method of claim 1, wherein the mixed population of cells is derived from pancreatic cell culture.

20. The method of claim 1, wherein the pancreatic Islet cells or progenitor or stem cells thereof are human.

21. The method of claim 15, wherein the pancreatic Islet cells or progenitor or stem cells thereof are of adult origin.

22. The method of claim 15, wherein the pancreatic Islet cells or progenitor or stem cells thereof are of fetal origin.

23. An enriched or purified preparation of isolated human pancreatic Islet cells or progenitor or stem cells.

24. The enriched or purified preparation of claim 23, wherein the cells are of adult origin.

25. The enriched or purified preparation of claim 23, wherein the cells are of fetal origin.

26. A method of treating a diabetic condition comprising:

providing the enriched or purified preparation of pancreatic Islet progenitor or stem cells or Islet cells differentiated therefrom and

transplanting the cells into a subject under conditions effective to treat a diabetic condition.

27. A method of treating a diabetic condition according to claim 26, wherein the diabetic condition is Diabetes Mellitus Type 1.

28. A method of treating a diabetic condition according to claim 26, wherein the diabetic condition is Diabetes Mellitus Type 2.

29. A method of treating a diabetic condition according to claim 26, wherein the cells are human.

30. A method of treating a diabetic condition according to claim 29, wherein the cells are of adult origin.

31. A method of treating a diabetic condition according to claim 29, wherein the cells are of fetal origin.

32. A method of treating a diabetic condition according to claim 29, wherein the cells produce insulin.

33. A method of treating a diabetic condition according to claim 29, wherein the cells produce glucagon.