DNER-MEDIATED CELL PURIFICATION OF PANCREATIC ENDOCRINE PRE-PROGENITOR CELLS AND THEIR PROGENY

• US 20110020297A1
• Filed: 04/03/2009
• Published: 01/27/2011
• Est. Priority Date: 04/03/2008
• Status: Active Grant

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Gradwohl, Gerard, Mellitzer, Georg, Madsen, Ole Dragsbaek, Hald, Jacob
• Granted Patent: US 8,399,251 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: G01N 33/56966 Animal cells

Abstract

The invention relates to a method of identifying and obtaining a culture cells comprising cells selected from the group consisting of endocrine pre-progenitor cells, endocrine progenitor cells, early endocrine cells, and/or fully differentiated endocrine cells. Also contemplated is a method of expanding the number of such cells as well as sorting such cells. In some aspects the invention further relates to a selective cell surface marker, DNER, that permits the selection of a unique subset of cells with endocrine pre-progenitor, endocrine progenitor, early endocrine, and/or fully differentiated endocrine phenotype. In some aspects the selective cell surface marker is selected from the group consisting of DNER, DISP2, SEZ6L2, LRP1 1 and SLC30A8. Furthermore, the invention relates to isolated cells selected from such cells and compositions thereof.

Claims

1. A method of identification of cells that have the potential to differentiate into pancreatic cells, the method comprising contacting a cell population comprising cells selected from the group consisting of endocrine pre-progenitor cells, endocrine progenitor cells, early endocrine cells, and/or fully differentiated endocrine cells with a DNER binding reagent wherein the cells that bind to the DNER reagent (DNER-positive cells) have the potential to differentiate into pancreatic cells.

2. A method of obtaining a culture of cells according to claim 1, the method further comprising:

separating the cells that bind the DNER binding reagent from cells that do not bind the DNER binding reagent wherein the cells that bind to the DNER reagent (DNER-positive cells) have the potential to differentiate into pancreatic cells.

3. A method of obtaining a culture of pancreatic cells, the method comprising:

obtaining cells that bind the DNER reagent (DNER-positive cells) according to the method of claim 2 and then subsequently culturing the DNER-positive cells under conditions which facilitate differentiation of the DNER-positive cells into pancreatic cells.

4. A method of expanding the number of cells in a cell population that have the potential to differentiate into pancreatic cells, the method comprising:

obtaining cells that bind the DNER reagent (DNER-positive cells) according to claim 2 and then subsequently culturing the obtained cells under conditions which facilitate expansion of the DNER-positive cell type(s) obtained.

5. A method of expanding the number of pancreatic cells in a cell population, the method comprising:

obtaining cells expanded according to claim 4 and then subsequently culturing the obtained cells under conditions which facilitates differentiation of the cells into pancreatic cells.

6. A method of providing pancreatic endocrine function to a mammal deficient in its production of at least one pancreatic hormone, the method comprising the steps of:

implanting into the mammal the cells obtained by the method according to claim 3 in an amount sufficient to produce a measurable amount of at least one pancreatic hormone in the mammal.

9. The method according to claim 1, wherein one or more additional binding reagents are used in combination with the DNER binding reagent either simultaneously or sequentially.

10. The method according to claim 9, wherein an additional binding reagent is selected from the group consisting of DDR1, prominin 1 (also known as CD133), CD49f, DISP2, SEZ6L2, LRP11 and SLC30A8 binding reagents.

11. The method according to claim 1, wherein the DNER positive cells are differentiated further into insulin producing cells, optionally together with cells differentiated further into cells selected from the group consisting of glucagon, somatostatin, pancreatic polypeptide, and/or ghrelin producing cells.

12. An isolated cell obtained by a method according to claim 1.

13. A composition comprising cells obtained by a method according to claim 1.

7. A method of quantifying DNER positive pancreatic cells by a) contacting the cells with a DNER binding reagent;

and b) determining the quantity of cells that bind the DNER reagent (DNER positive cells).

8. (canceled)

14-15. -15. (canceled)