Method of expanding hepatic precursor cells

• US 5,576,207 A
• Filed: 06/24/1994
• Issued: 11/19/1996
• Est. Priority Date: 08/07/1991
• Status: Expired due to Term

• Current Assignee: Albert Einstein College of Medicine of Yeshiva University
• Original Assignee: Albert Einstein College of Medicine of Yeshiva University
• Inventors: Reid, Lola M., Ochs, Andreas, Agelli, Maria
• Primary Examiner(s): Walsh, Stephen G.
• Assistant Examiner(s): KEMMERER, ELIZABETH
• Application Number:
  US08/265,696
• Time in Patent Office: 879 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23
• US Class Current: 435/378
• 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: C12N 2500/20 Transition metals
• CPC Class Codes: C12N 2500/22 Zinc; Zn chelators
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/305 Growth hormone [GH], aka. somatotropin
• CPC Class Codes: C12N 2501/315 Prolactin
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/395 Thyroid hormones
• CPC Class Codes: C12N 2502/02 embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 2533/70 Polysaccharides
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0672 Stem cells; Progenitor cells; Precursor cells; Oval cells

Abstract

A composition which comprises an animal cell population, and which contains immature animal cells. The immature animal cells are characterized by expression of alpha-fetoprotein or lack of essential expression of alpha-fetoprotein and albumin, and at least a portion of said immature animal cells or at least a portion of the progeny of said immature cells is capable of differentiating into cells which express albumin. The cell population is cultured under conditions which result in expansion of the cells. Expansion of the cells may be achieved by culturing the cells in the presence of an extracellular matrix and liver stromal cells; and preferably in the presence of growth factors. Such cells may be used for liver transplantation, artificial livers, and for toxicology and pharmacology studies. Such cells may also be genetically engineered to express proteins or polypeptides of interest.

Claims

1. A method of expanding hepatic precursor cells comprising culturing immature liver cells which contain at least a population of hepatic precursor cells in serum-free culture medium, an extracellular matrix and liver stromal cells such that hepatic precursor cells contained within the cell culture are expanded.

2. The method of claim 1 wherein the hepatic precursor cells are expanded at least three fold.

3. The method of claim 1 wherein the extracellular matrix is formed from a material selected from the group consisting of collagen, fibronectin, laminin, proteoglycan and combinations thereof.

4. The method of claim 5 wherein the collagen is Type IV.

5. The method of claim 1 wherein the liver stromal cells are selected from the group consisting of embryonic liver stromal cells and fetal liver stromal cells.

6. The method of claim 1 wherein the culture further includes a growth factor.

7. A method of obtaining hepatic precursor cells comprising:

(a) culturing immature liver cells which contain at least a population of hepatic precursor cells in serum-free culture medium, an extracellular matrix and liver stromal cells such that hepatic precursor cells contained within the cell culture are expanded; and

(b) separating the expanded hepatic precursor cells from the culture medium so as to obtain hepatic precursor cells.

8. The method of claim 7 wherein the hepatic precursor cells are expanded at least three fold.

9. The method of claim 7 wherein the extracellular matrix is formed from a material selected from the group consisting of collagen, fibronectin, laminin and combinations thereof.

10. The method of claim 7 wherein the collagen is Type IV.

11. The method of claim 7 wherein the liver stromal cells are selected from the group consisting of embryonic liver stromal cells and fetal liver stromal cells.

12. The method of claim 7 wherein the culture further includes a growth factor.

13. A method of obtaining hepatic precursor cells comprising:

(a) culturing immature liver cells which contain at least a population of hepatic precursor cells in serum-free culture medium, an extracellular matrix, at least one growth factor, and liver stromal cells such that hepatic precursor cells contained within the cell culture are expanded; and

(b) separating the expanded hepatic precursor cells from the culture medium so as to obtain hepatic precursor cells.