ENDODERMAL PROGENITOR CELLS

• US 20100150876A1
• Filed: 11/26/2007
• Published: 06/17/2010
• Est. Priority Date: 11/24/2006
• Status: Active Grant

• Current Assignee: Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Chase, Lucas, Verfaillie, Catherine
• Granted Patent: US 9,005,964 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A01K 2267/0325 Animal model for autoimmune diseases
• 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: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• CPC Class Codes: C12N 5/0678 Stem cells; Progenitor cells; Precursor cells

Abstract

The present invention relates to endodermal progenitor cells and methods of isolation, culture, differentiation and use thereof.

Claims

1. A method of obtaining endodermal progenitor cells comprising:

(a) culturing a cell population obtained from a tissue to induce cluster formation in culture medium;

(b) culturing the clusters in culture medium; and

(c) isolating endodermal progenitor cells,

wherein a function of a cell in the tissue is not reduced by application of an exogenous stressor prior to harvesting the tissue.

2. The method of claim 1, wherein the tissue is endodermal tissue.

3. The method of claim 1, wherein the tissue is derived from liver, stomach, intestine, pancreas, lung, colon, bladder or thyroid.

4. The method of claim 1, wherein the tissue is pancreas.

5. The method of claim 1, wherein the clusters are isolated prior to culturing in step (b).

6. The method of claim 1, wherein said clusters of step (b) are cultured with one more or attachment factors.

7. The method of claim 6, wherein the one or more attachment factors comprise one or more of collagen type I, collagen type II, collagen type IV, fibronectin, chondroitin sulfate, vitronectin, thrombospondin, or matrigel.

8. The method of claim 1, wherein the culture medium of step (b) is changed every two to three days for at least about 7 days.

9. The method of claim 8, wherein after about 7 days the culture medium of step (b) is not changed prior to isolating the endodermal progenitor cells.

10. The method of claim 1, wherein the culture medium of step (a) and/or (b) comprises serum.

11. The method of claim 10, wherein the serum content is about 0.5% to about 5%.

12. The method of claim 1, wherein the culture media of step (a) and/or (b) comprises one or more of EGF, LIF, PDGF, or basic FGF.

13. The method of claim 1, wherein the cell population is subjected to continuous density gradient centrifugation prior to step (a).

14. The method of claim 1, wherein the tissue is disassociated prior to step (a).

15. The method of claim 14, wherein the tissue is disassociated enzymatically.

16. The method of claim 15, wherein the enzyme comprises one or more of collagenase, trypsin, dispase I, hyaluronidase, thermolysin, neutral protease, liberase RI, DNase I, pancreatin, or pronase.

17. The method of claim 1, wherein the tissue is obtained from a mammal.

18. The method of claim 17, wherein the mammal is a human, swine, mouse or rat.

19. An endodermal progenitor cell prepared by the method of claim 1.

20. The endodermal progenitor cell of claim 19, wherein the progenitor cell originates in bone marrow.

21. A composition comprising a population of endodermal progenitor cells of claim 20 and a pharmaceutically acceptable carrier and/or culture medium.

22. The method of claim 1 further comprising differentiating the endodermal progenitor cells to yield an endodermal cell type.

23. The method of claim 22, wherein the differentiated endodermal cell type is selected from the group consisting of pancreatic, islet, intestinal, thyroid, lung, colon, bladder and liver cell types.

24. The method of claim 22, wherein the endodermal progenitor cells are differentiated in the presence of one or more differentiation factors comprising β

-cellulin, GLP-1, HGF, FGF, EGF, KGF, nicotinamide, TGF-α

, TGF-β

, activin, cyclopamin, BMP4, an SHH antibody, FGF, EGF, Oncostatin M, dexamethasone, exendin4, GDF11, nicotinamide or a combination thereof.

25. A method for providing an endodermal cell type to a subject in need thereof, comprising administering to said subject the endodermal progenitor cells produced by the method of claim 1 or differentiated progeny derived therefrom in an amount effective to provide an endodermal cell type to the subject.

26. The method of claim 25, wherein the endodermal progenitor cells or differentiated progeny derived therefrom are administered by systemic or local injection.

27-30. -30. (canceled)