Liver stem cell

• US 6,872,389 B1
• Filed: 07/08/1999
• Issued: 03/29/2005
• Est. Priority Date: 07/08/1999
• Status: Expired due to Term

• Current Assignee: Rhode Island Hospital
• Original Assignee: Rhode Island Hospital
• Inventors: Faris, Ronald A.
• Primary Examiner(s): WOITACH, JOSEPH T
• Application Number:
  US09/743,544
• Time in Patent Office: 2,091 Days
• Field of Search: 435/325, 435/1.1, 424/93.7, 424/93.1
• US Class Current: 424/93.7
• CPC Class Codes: C12N 2501/58 Adhesion molecules, e.g. ICAM, VCAM, CD18 (ligand), CD11 (ligand), CD49 (ligand)
• CPC Class Codes: C12N 5/0672 Stem cells; Progenitor cells; Precursor cells; Oval cells

Abstract

The invention provides methods of isolating a primary liver stem cell by identifying a cell doublet containing a hepatocytes and the stem cell and isolating the doublet from liver tissue.

Claims

1. A method of obtaining a population of liver cell clusters comprising, isolating a liver cell cluster from liver tissue to yield a population of clusters, wherein said cluster consists essentially of a total of 2 to 5 cells at least one of which is a stem cell associated with a hepatocyte and wherein said stem cell comprises an OV6 antigen and lacks an OC2 antigen.

2. The method of claim 1, further comprising enriching said liver tissue for periportal hepatocytes associated with the biliary tree, wherein said enriching occurs prior to said isolating said liver cell cluster from liver tissue.

3. The method of claim 1, wherein said liver cell cluster is a cell doublet.

4. The method of claim 1, further comprising enriching said population of liver cell clusters for liver cell clusters from the canal of Hering of an adult liver, said enriching comprising identifying and isolating liver cell clusters expressing a bile duct cell marker.

5. The method of claim 4, wherein said bile duct cell marker is cytokeratin 19 (CK 19).

6. The method of claim 1, further comprising selecting for expression of desmoplakin, wherein said selecting for expression of desmoplakin occurs after said isolating said liver cell cluster from liver tissue.

7. The method of claim 1, further comprising selecting for expression of OV6, wherein said selecting for expression of OV6 occurs after said isolating said liver cell cluster from liver tissue.

8. The method of claim 1, further comprising selecting for a cell which expresses an antigen selected from the group consisting of laminin, desmoplakin I, CCAM, CEA, dipeptidyl peptidase-4, γ

GT, VLA-2, VLA-3, VLA-5, and VLA-6.

9. The method of claim 1, wherein said liver tissue is fetal liver tissue.

10. The method of claim 1, wherein said liver tissue is pediatric liver tissue.

11. The method of claim 1, wherein said liver tissue is adult liver tissue.

12. The method of claim 1, wherein said liver tissue is obtained from a mouse, rat, dog, baboon, or pig.

13. The method of claim 1, wherein said liver tissue is obtained from a human.

14. The method of claim 1, wherein said liver tissue is obtained from a living or deceased donor.

15. The method of claim 1, wherein said liver tissue is normal liver tissue.

16. The method of claim 1, wherein said cluster is isolated from said liver tissue prior to a mechanical injury or exposure to a carcinogen.

17. The method of claim 1, wherein said population of clusters comprises at least 60% doublets.

18. The method of claim 1, wherein said population of clusters comprises at least 90% doublets.

19. The method of claim 1, wherein said population of clusters comprises at least 99% doublets.

20. The method of claim 1, further comprising dissociating said stem cell from said hepatocyte and removing said hepatocyte to yield a sample of substantially pure liver stem cells.

21. The method of claim 1, wherein said isolating a liver cell cluster from liver tissue comprises identifying a desmosomal junction between said stem cell and said hepatocyte.

22. The method of claim 1, wherein said cluster consists essentially of a total of 3 to 5 cells at least two of which are stem cells associated with a hepatocyte.

23. The method of claim 22, wherein said isolating a liver cell cluster from liver tissue comprises identifying a desmosomal junction between at least one of said stems cells and said hepatocyte.

24. The method of claim 1, wherein said liver cell cluster is a cell triplet.