Cardiomyocyte medium with dialyzed serum

• US 9,365,826 B2
• Filed: 06/20/2011
• Issued: 06/14/2016
• Est. Priority Date: 06/18/2010
• Status: Active Grant

• Current Assignee: FUJIFILM Cellular Dynamics, Inc. (Fujifilm Holdings Corporation)
• Original Assignee: Cellular Dynamics International, Inc. (Fujifilm Holdings Corporation)
• Inventors: Meyer, Nathan, Swanson, Brad, Fiene, Steve
• Primary Examiner(s): Kim, Taeyoon
• Application Number:
  US13/164,461
• Publication Number:
  US 20110312090A1
• Time in Patent Office: 1,821 Days
• Field of Search: 435/325, 435/366, 435/374, 435/14
• US Class Current: 1/1
• CPC Class Codes: C12N 2500/00 Specific components of cell culture medium
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells

Abstract

Methods and composition for maintenance of cardiomyocytes are provided. For example, in certain aspects methods including culturing the cardiomyocytes in a medium essentially free of serum or containing dialyzed serum to maintain long-term purity. In further aspects, methods for modulation of cardiomyocytes may be provided.

Claims

1. A composition comprising a cell population of at least 90% cardiomyocytes and a medium that is essentially free of glucose, wherein said cardiomyocytes are prepared from human induced pluripotent stem (iPS) cells and further wherein said cells have been cultured in said medium for at least 7 days.

2. The composition of claim 1, wherein the medium comprises dialyzed serum.

3. The composition of claim 2, wherein the medium comprises dialyzed serum in a range of about 1% to about 75%.

4. The composition of claim 3, wherein the medium comprises dialyzed serum of at least 10%.

5. The composition of claim 4, wherein the medium comprises dialyzed serum of at least 25%.

6. The composition of claim 4, wherein the medium comprises dialyzed serum of about 55%.

7. The composition of claim 2, wherein said serum has been treated to remove low molecular weight molecules having a molecular weight of at most 10 kD.

8. The composition of claim 2, wherein said serum is essentially free of low molecular weight molecules having a molecular weight of at most 10 kD.

9. The composition of claim 1, wherein the medium does not comprise glucose.

10. The composition of claim 1, wherein the medium comprises about 1 to 20 mM galactose.

11. The composition of claim 1, wherein the medium comprises about 0.1 to 10 mM pyruvate or pyruvic acid.

12. The composition of claim 1, wherein the composition comprises a cell population of at least 95% cardiomyocytes.

13. The composition of claim 1, wherein the cardiomyocytes express at least one selectable or screenable transgene under the control of a cardiomyocyte-specific promoter.

14. The composition of claim 1, wherein the population is essentially free of fibroblasts or undifferentiated pluripotent stem cells.

15. The composition of claim 1, wherein the cardiomyocytes are cryopreserved cardiomyocytes.