Liver stem cell

• US 20060019387A1
• Filed: 01/19/2005
• Published: 01/26/2006
• Est. Priority Date: 07/08/1999
• Status: Active Grant

• Current Assignee: Rhode Island Hospital
• Original Assignee: Rhode Island Hospital
• Inventors: Faris, Ronald A.
• Granted Patent: US 7,935,528 B2
• Time in Patent Office: Days
• US Class Current: 435/370
• 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 a primary liver stem cell and a cell doublet consisting of a hepatocyte and the stem cell, both of which are derived from normal liver tissue. Methods of isolating the cells, genetically altering the cells, and using the cells for transplantation are also within the invention.

Claims

1. A primary liver stem cell, wherein said stem cell is

(a) obtained from normal liver tissue, and

(b) isolated from an isolated liver cell cluster comprising a hepatocyte and said stem cell,

wherein said stem cell comprises a DNA encoding a heterologous protein.

2. The liver stem cell of claim 1, wherein said heterologous protein is selected from the group consisting of ornithine transcarbamylase, arginosuccinate synthetase, glutamine synthetase, glycogen synthetase, glucose-6-phosphatase, succinate dehydrogenase, glucokinase, pyruvate kinase, acetyl CoA carboxylase, fatty acid synthetase, alanine aminotransferase, glutamate dehydrogenase, ferritin, LDL receptor, alcohol dehydrogenase, albumin, transferrin, complement component C3, α

2-macroglobulin, fibrinogen, Factor XIII;

C, Factor IX, and α

1-antitrypsin.

3. The liver stem cell of claim 1, wherein said hepatocyte and said stem cell are joined by a desmosomal junction.

4. The liver stem cell of claim 1, wherein said stem cell is a pre-oval cell.

5. The liver stem cell of claim 1, wherein said stem cell expresses OV6.

6. The liver stem cell of claim 1, wherein said stem cell expresses a bile duct cell marker.

7. The liver stem cell of claim 6, wherein said bile ductal cell marker is a cytokeratin.

8. The liver stem cell of claim 7, wherein said cytokeratin is cytokeratin 19.

9. The liver stem cell of claim 3, wherein said stem cell expresses desmoplakin.

10. An isolated liver cell cluster comprising a liver stem cell and a hepatocyte, wherein said stem cell comprises a DNA encoding a heterologous protein.

11. The liver cell cluster of claim 10, wherein said heterologous protein is selected from the group consisting of ornithine transcarbamylase, arginosuccinate synthetase, glutamine synthetase, glycogen synthetase, glucose-6-phosphatase, succinate dehydrogenase, glucokinase, pyruvate kinase, acetyl CoA carboxylase, fatty acid synthetase, alanine aminotransferase, glutamate dehydrogenase, ferritin, low density lipoprotein (LDL) receptor, alcohol dehydrogenase, albumin, transferrin, complement component C3, α

2-macroglobulin, fibrinogen, Factor XIII;

C, Factor IX, and α

1-antitrypsin.

12. The liver cell cluster of claim 10, wherein said liver cell cluster is a cell doublet.

13. The liver cell cluster of claim 10, wherein said hepatocyte and said stem cell are joined by a desmosomal junction.

14. The liver cell cluster of claim 10, wherein said stem cell is a pre-oval cell.

15. The liver cell cluster of claim 10, wherein said stem cell expresses OV6.

16. The liver cell cluster of claim 10, wherein said stem cell expresses a bile duct cell marker.

17. The liver cell cluster of claim 16, wherein said bile ductal cell marker is a cytokeratin.

18. The liver cell cluster of claim 17, wherein said cytokeratin is cytokeratin 19.

19. The liver cell cluster of claim 13, wherein said stem cell expresses desmoplakin.