Markers for identification and isolation of pancreatic islet α and β progenitors

• US 6,753,153 B2
• Filed: 12/13/2000
• Issued: 06/22/2004
• Est. Priority Date: 12/13/1999
• Status: Expired due to Fees

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Kritzik, Marcie, Sarvetnick, Nora
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Padmanabhan, Kartic
• Application Number:
  US09/736,911
• Publication Number:
  US 20020106703A1
• Time in Patent Office: 1,287 Days
• Field of Search: 435/2, 435/4, 435/7.21, 435/7.23, 435/7.24, 435/7.25, 435/371, 435/377, 435/373, 435/378, 435/325, 436/519, 436/546, 436/548, 436/63, 436/64, 436/172, 436/177
• US Class Current: 435/7.21
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/025 Animal producing cells or organs for transplantation
• CPC Class Codes: A01K 2267/0325 Animal model for autoimmune diseases
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• 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: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

The differential expression of marker proteins in a targeted population provides a means of identifying and isolating cells. A population of cells associated with the regeneration of pancreatic islets is shown to express certain proteins, including the cell surface proteins ErbB2, ErbB3, and ErbB4; and the nuclear protein Msx-2. Populations of isolated pancreatic islet progenitor cells find use in screening assays, to characterize genes involved in islet development and regulation, and in transplantation to provide a recipient with pancreatic islet functions.

Claims

1. A method of identifying mammalian pancreatic islet progenitor cells, the method comprising:

contacting a population of normal fetal or neonatal mammalian pancreatic cells with marker ErbB2 specific binding members comprising a detectable label ErbB2; and

detecting those cells that bind to said ErbB2 specific binding members;

wherein cells that bind to said ErbB2 specific binding members are identified as pancreatic islet progenitor cells.

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

3. The method of claim 1, wherein said pancreatic cells are mouse.

4. The method of claim 1, wherein said marker ErbB2 specific binding member is an antibody.

5. The method of claim 1, wherein said detecting step comprises detection of said label by flow cytometry.

6. The method of claim 5, further comprising the step of:

separating the cells in said population based on binding to said ErbB2 specific binding members to provide a purified population of pancreatic progenitor cells.

7. An isolated population of pancreatic islet progenitor cells, wherein said cells are derived from normal fetal or neonatal pancreatic ducts, and are characterized as expressing ErbB2.

8. The isolated cell population of claim 7, wherein said cells are human cells.

9. The isolated cell population of claim 7, wherein said cells are mouse cells.

10. An in vitro cell culture, comprising:

a cell population according to claim 7; and

cell culture medium.

11. A method of identifying mammalian pancreatic islet progenitor cells, the method comprising:

contacting a population of normal fetal or neonatal mammalian pancreatic cells with ErbB3-specific binding members comprising a detectable label; and

detecting those cells that bind to said ErbB3-specific binding members;

wherein cells that bind to said ErbB3-specific binding members are identified as pancreatic islet progenitor cells.

12. The method of claim 11, wherein said cells are human cells.

13. The method of claim 11, wherein said cells are mouse cells.

14. The method of claim 11, wherein said ErbB3-specific binding member is an antibody.

15. The method of claim 11 wherein said detecting step comprises detection of said label by flow cytometry.

16. The method of claim 11, further comprising the step of:

separating the cells in said population based on binding to said ErbB3-specific binding members to provide a purified population of pancreatic progenitor cells.

17. An isolated population of pancreatic islet progenitor cells, wherein said cells are derived from normal fetal or neonatal mammalian pancreatic cells, and are characterized as expressing ErbB3.

18. The isolated cell population of claim 17, wherein said pancreatic islet progenitor cells are progenitors for insulin producing beta cells.

19. The isolated cell population of claim 17, wherein said cells are further characterized as lacking detectable production of insulin.

27. The isolated cell population of claim 17, wherein said pancreatic islet progenitor cells are progenitors for glucagon producing alpha cells.

20. A method of identifying mammalian pancreatic islet progenitor cells, the method comprising:

contacting a population of normal fetal or neonatal mammalian pancreatic cells with ErbB4-specific binding members comprising a detectable label; and

detecting those cells that bind to said ErbB4-specific binding members;

wherein cells that bind to said ErbB4-specific binding members and that lack expression of glucagon or insulin are identified as pancreatic islet progenitor cells.

21. The method of claim 20, wherein said cells are human cells.

22. The method of claim 20, wherein said cells are mouse cells.

23. The method of claim 20, wherein said ErbB4-specific binding member is an antibody.

24. The method of claim 20, wherein said detecting step comprises detection of said label by flow cytometry.

25. The method of claim 20, further comprising the step of:

separating the cells in said population based on binding to said ErbB4-specific binding members to provide a purified population of pancreatic progenitor cells.

26. An isolated population of pancreatic islet progenitor cells, wherein said cells are derived from normal fetal or neonatal mammalian pancreatic cells, and are characterized as expressing ErbB4 and lacking detectable production of insulin or glucagon.