FORMULATIONS AND METHODS FOR CULTURING STEM CELLS

• US 20100081200A1
• Filed: 12/07/2009
• Published: 04/01/2010
• Est. Priority Date: 06/05/2007
• Status: Abandoned Application

• Current Assignee: Heli Skottman, Kristiina Rajala, Marjo-Riitta Suuronen, Outi Hovatta
• Original Assignee: Heli Skottman, Kristiina Rajala, Marjo-Riitta Suuronen, Outi Hovatta
• Inventors: Skottman, Heli, Hovatta, Outi, Rajala, Kristiina, Suuronen, Marjo-Riitta
• Application Number:
  US12/632,057
• Publication Number:
  US 20100081200A1
• Time in Patent Office: Days
• US Class Current: 435/377
• CPC Class Codes: C12N 2500/20 Transition metals
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/60 Buffer, e.g. pH regulation, osmotic pressure
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The present invention relates to a serum replacement formulation and to a culture medium suitable for the derivation, maintenance and differentiation of stem cells.

Claims

1. A xeno-free serum replacement comprising at least one fatty acid selected from a group consisting of conjugated linoleic acid and eicosapentaenoic acid.

2. The serum replacement according to claim 1, further comprising Activin A.

3. The serum replacement according to claim 1, further comprising retinol.

4. The serum replacement according to claim 1 further comprising stearic acid.

5. The serum replacement according to claim 1, wherein, the concentration of conjugated linoleic acid (CLA) is such that a final culture medium, which is a basal medium supplemented with said serum replacement, comprises from about 0.5 mg/l to about 5 mg/l CLA, and the concentration of eicosapentaenoic acid (EPA) is such that the final culture medium comprises from about 1 mg/l to about 10 mg/l EPA.

6. The serum replacement according to claim 2, wherein the concentration of Activin A is such that a final culture medium, which is a basal medium supplemented with said serum replacement, comprises from about 0.001 mg/l to about 0.02 mg/l Activin A.

7. The serum replacement according to claim 3, wherein the concentration of retinol is such that a final culture medium, which is a basal medium supplemented with said serum replacement, comprises from about 0.25 mg/l to about 0.5 mg/l retinol.

8. The serum replacement according to claim 4, wherein the concentration of stearic acid is such that a final culture medium, which is a basal medium supplemented with said serum replacement, comprises from about 0.5 mg/l to about 5 mg/l stearic acid.

9. A xeno-free cell culture medium comprising a basal medium and the serum replacement according to claim 1.

10. A method for initiating a new stem cell line in vitro, comprising

a) providing isolated cells of desired origin,

b) contacting said cells with the xeno-free medium according to claim 9, and

c) cultivating said cells under conditions suitable for stem cell culture.

11. The method according to claim 10, wherein said isolated cells are of embryonic, adult somatic, or mesenchymal origin.

12. A method for culturing stem cells, comprising

a) contacting said stem cells with the xeno-free medium according to claim 9, and

b) cultivating said cells under conditions suitable for stem cell culture.

13. The method according to claim 11, wherein said cultivation is performed on a feeder cell layer.

14. A method for differentiating stem cells, comprising

a) contacting said stem cells with the xeno-free medium according to claim 9 supplemented with a differentiating agent, and

b) cultivating said cells under conditions suitable for differentiation of stem cells.

15. The serum replacement according to claim 2, further comprising retinol.

16. The serum replacement according to claim 15, wherein the concentration of retinol is such that a final culture medium, which is a basal medium supplemented with said serum replacement, comprises from about 0.25 mg/l to about 0.5 mg/l retinol.

17. The method according to claim 12, wherein said cultivation is performed on a feeder cell layer.