Cell culture medium for enhanced hepatocyte function

Patent metadata

Field	Value
Publication number	US-10876096-B2
Application number	US-201314647304-A
Country	US
Kind code	B2
Filing date	Nov 25, 2013
Priority date	Nov 28, 2012
Publication date	Dec 29, 2020
Grant date	Dec 29, 2020

Abstract

Cell culture compositions containing LFM-A13 or a structurally related compound can enhance global hepatic function. For example, LFM-A13 is shown to enhance levels of a broad variety of drug metabolism enzymes, including CYP enzymes, and other hepatic enzymes. LFM-A13 is also shown to promote differentiation of stem cells into hepatocytes. LFM-A13 and structurally related compounds can be used in cell culture to enhance global drug metabolism of liver cells for enhanced in vitro study the effects of drug metabolism on other candidate drug compounds.

First claim

What is claimed is:

1. A composition for producing metabolites of a candidate compound in a culture of liver cells, the composition comprising:
a cell culture medium;
the liver cells;
the candidate compound; and
a compound of Formula:

wherein the LFM-A13 is present in the composition in an amount effective to increase levels of more than one cytochrome P450 monooxygenase (CYP) enzyme by greater than two-fold in the culture of the liver cells relative to liver cells cultured in the same composition but lacking the LFM-A13, and
wherein the more than one CYP enzyme is chosen from CYP3A4, CYP1A2, or CYP2B6.

2. The composition according to claim 1 , wherein the LFM-A13 is present in the composition at a concentration from about 0.1 micromolar to about 50 micromolar.

3. The composition according to claim 1 , wherein the LFM-A13 is present in the composition at a concentration from about 1 micromolar to about 20 micromolar.

4. The composition according to claim 1 , wherein the cell culture medium comprises serum.

5. The composition according to claim 4 , wherein the cell culture medium comprises nutrients capable of supporting cell growth including amino acids, energy sources, and salts, and wherein:
the energy sources comprise glucose, and
the salts provide at least one inorganic ion chosen from Na + , K + , Ca 2+ , Cu 2+ , Zn 2+ , and Co + .

6. The composition according to claim 1 , wherein the cell culture medium is free from serum.

7. The composition according to claim 1 , wherein the liver cells comprise primary hepatocytes.

8. The composition according to claim 1 , wherein the liver cells comprise immortalized cells derived from a hepatocyte.

9. The composition according to claim 1 , wherein the liver cells comprise a cell line derived from a hepatocyte.

10. The composition according to claim 1 , wherein the liver cells comprise a cell line derived from an embryonic stem cell.

11. The composition according to claim 1 , wherein the LFM-A13 is present in the composition at a concentration of about 10 micromolar.

12. The composition according to claim 1 , wherein the more than one CYP enzyme is CYP3A4 and CYP1A2.

13. The composition according to claim 1 , wherein:
the more than one CYP enzyme is CYP3A4 and CYP1A2; and
the LFM-A13 is present in the composition in an amount effective to increase levels of the more than one cytochrome P450 monooxygenase (CYP) enzyme by at least three-fold in the culture of the liver cells relative to liver cells cultured in the same composition but lacking the LFM-A13.

14. A method for enhancing the metabolic function of liver cells in culture, comprising:
culturing the liver cells in the composition according to claim 1 , wherein metabolic function of the liver cells is enhanced by the LFM-A13 present in the composition in the amount effective to increase the levels of the more than one CYP enzyme.

15. The method according to claim 14 , wherein the liver cells comprise primary hepatocytes.

16. The method according to claim 14 , wherein the liver cells comprise immortalized cells derived from a hepatocyte or from an embryonic stem cell.

17. The method according to claim 14 , wherein the liver cells comprise a cell line.

18. The method according to claim 17 , wherein the cell line is derived from a hepatocyte or from an embryonic stem cell.

19. A method for identifying metabolites of a candidate compound, comprising:
culturing the liver cells in the composition according to claim 1 , wherein the cultured cells are contacted with the candidate compound; and
identifying the metabolites of the candidate compound.

Assignees

• Corning Inc
• Faris Ronald Allen
• Hong Yulong
• Lahiri Joydeep
• Liu Jin
• Weeks Olivia Grace

Inventors

• Faris Ronald Allen
• Hong Yulong
• Lahiri Joydeep
• Liu Jin
• Weeks Olivia Grace

Classifications

• C12N2501/727

  Kinases (EC 2.7.) · CPC title

• C12N2533/54

  Collagen; Gelatin · CPC title

• C12N2503/02

  Drug screening · CPC title

• C12Q2600/158

  Expression markers · CPC title

• C12N5/067 Primary

  Hepatocytes · CPC title