DDR1-Mediated Cell Purification of Pancreatic Endocrine Cells Through Their Progenitors

• US 20110014160A1
• Filed: 12/19/2008
• Published: 01/20/2011
• Est. Priority Date: 12/28/2007
• Status: Active Grant

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Madsen, Ole Dragsbaek, Serup, Palle, Hald, Jacob
• Granted Patent: US 8,551,779 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• 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 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: G01N 33/507 Pancreatic cells

Abstract

The invention relates to a method of identifying, obtaining and/or quantifying a culture of pancreatic cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, early endocrine cells, and fully differentiated endocrine cells. Also contemplated is a method of expanding the numbers of such cells as well as sorting such cells. The invention further relates to a selective cell surface marker, DDR1, that permits the selection of a unique subset of cells with pancreatic endocrine progenitor phenotype. Furthermore, the invention relates to isolated cells selected from such cells and compositions thereof.

Claims

1. A method of identification of pancreatic cells comprising cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, and early endocrine cells, the method comprising contacting a cell population comprising pancreatic cells with a DDR1 binding reagent.

9. An isolated cell selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, early endocrine cells obtained by a method as defined in claim 1 and fully differentiated endocrine cells obtained by a method as defined in claim 3.

10. An isolated endocrine progenitor cell obtained by a method as defined in claim 1.

12. A composition comprising isolated cells selected from one or more cells from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, early endocrine cells obtained by a method as defined in claim 1 and fully differentiated endocrine cells.

2. A method of obtaining a culture of pancreatic cells comprising cells selected from the group consisting of ductal/endocrine progenitor cells, pancreatic endocrine progenitor cells, and early endocrine cells comprising:

contacting a cell population comprising pancreatic cells with a DDR1 binding reagent and separating the cells that binds the DDR1 binding reagent in a fraction of DDR1 positive cells from cells that do not bind the DDR1 binding reagent.

3. The method of obtaining a culture of pancreatic cells comprising cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, and early endocrine cells comprising:

obtaining cells purified according to the method of claim 2 and then subsequently culturing the obtained cells under conditions which facilitate their differentiation into pancreatic cells comprising cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, early endocrine cells, and fully differentiated endocrine cells.

4. The method of expanding the numbers of pancreatic cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, and early endocrine cells comprising:

obtaining cells purified according to claim 2 and then subsequently culturing the obtained cells under conditions which facilitate expansion of the cell type(s) obtained.

5. The method of expanding the numbers of pancreatic cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, and early endocrine cells comprising:

obtaining cells purified and expanded according to claim 4 and then subsequently culturing the obtained cells under conditions which facilitates differentiation of the cells into pancreatic cells selected from the group consisting of ductal/endocrine progenitor cells, endocrine progenitor cells, early endocrine cells, and fully differentiated endocrine cells.

6. The method of providing pancreatic endocrine function to a mammal deficient in its production of at least one pancreatic hormone wherein cells are obtained by the method according to claim 5, the method further comprising the steps of:

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

11. An isolated fully differentiated endocrine cell obtained by a method as defined in claim 3.

7. A method of quantifying DDR1 positive cells by a) contacting the cells with a DDR1 binding reagent;

and b) determining the quantity of cells that exhibit DDR1 as a cell surface marker (DDR1 positive cells).

8. A method for the optimisation of an in vitro protocol, wherein the number of DDR1 ex-pressing cells (DDR1 positive cells) is periodically monitored.

13. (canceled)

14. (canceled)