Media for culturing stem cells

Patent metadata

Field	Value
Publication number	US-9404079-B2
Application number	US-201313909128-A
Country	US
Kind code	B2
Filing date	Jun 4, 2013
Priority date	Aug 29, 2005
Publication date	Aug 2, 2016
Grant date	Aug 2, 2016

Abstract

Well-defined, xeno-free culture media which comprise a TGF-beta isoform or the chimera formed between IL6 and the soluble IL6 receptor (IL6RIL6), which are capable of mainataining stem cells, and particularly, human embryonic stem cells, in an undifferentiated state are provided. Also provided are cell cultures comprising the culture media and the stem cells and methods of expanding and deriving embryonic stem cells in such well-defined, xeno-free culture media. In addition, the present invention provides methods of differentiating ESCs or EBs formed therefrom for the generation of lineage specific cells.

First claim

What is claimed is:

1. A method of expanding and maintaining human pluripotent stem cells in an undifferentiated state, the method comprising culturing the human pluripotent stem cells in a culture medium comprising a transforming growth factor beta 3 (TGFβ3) isoform— and factors that maintain and expand human pluripotent stem cells in an undifferentiated state, thereby expanding and maintaining the human pluripotent stem cells in the undifferentiated state.

2. The method of claim 1 , wherein said human pluripotent stem cells are human embryonic stem cells.

3. The method of claim 1 , wherein the human pluripotent stem cells are cultured in suspension.

4. The method of claim 1 , wherein the human pluripotent stem cells are cultured on a feeder-layer free matrix.

5. The method of claim 4 , wherein said feeder-layer free matrix is a fibronectin matrix.

6. The method of claim 1 , wherein the human pluripotent stem cells are cultured on feeder cells.

7. The method of claim 1 , wherein said TGFβ3 isoform is provided at a concentration of at least 0.5 ng/ml.

8. The method of claim 1 , wherein said TGFβ3 isoform is provided at a concentration of at least 2 ng/ml.

9. The method of claim 1 , wherein said culture medium further comprises basic fibroblast growth factor (bFGF).

10. The method of claim 9 , wherein said bFGF is provided at a concentration of at least 2 ng/ml.

11. The method of claim 1 , wherein said culture medium is serum-free.

12. The method of claim 1 , wherein said culture medium is devoid of animal contaminant.

13. The method of claim 3 , wherein said suspension is protein carrier-free.

Assignees

• Technion Res & Dev Foundation

Inventors

• Amit Michal
• Itskovitz-Eldor Joseph

Classifications

• C12N5/0056

  Xeno-free medium · CPC title

• C12N5/0603

  Embryonic cells (production of embryos, nuclear transfer A01K67/027); Embryoid bodies · CPC title

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title

• C12N5/0602

  Vertebrate cells · CPC title

• C12N5/0606 Primary

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title