Human foreskin fibroblast conditioned media for culturing ES cells

• US 7,790,455 B2
• Filed: 07/06/2007
• Issued: 09/07/2010
• Est. Priority Date: 10/07/2002
• Status: Active Grant

• Current Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD.
• Original Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD.
• Inventors: Itskovitz-Eldor, Joseph, Amit, Michal
• Primary Examiner(s): Crouch; Deborah
• Application Number:
  US11/822,514
• Publication Number:
  US 20070259426A1
• Time in Patent Office: 1,159 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: C12N 2500/00 Specific components of cell culture medium
• CPC Class Codes: C12N 2500/99 Serum-free medium
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0031 Serum-free culture media
• CPC Class Codes: C12N 5/0043 Medium free of human- or animal-derived components
• CPC Class Codes: C12N 5/0056 Xeno-free medium
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• 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
• CPC Class Codes: C12N 5/0611 Primordial germ cells, e.g. embryonic germ cells [EG]
• CPC Class Codes: C12N 5/0665 Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
• CPC Class Codes: C12N 5/0695 Stem cells; Progenitor cells; Precursor cells
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

A cell culture comprising human foreskin cells, the human foreskin cells being capable of maintaining stem cells in an undifferentiated state when co-cultured therewith.

Claims

1. A method of maintaining embryonic stem cells in an undifferentiated state comprising culturing the embryonic stem cells with a human foreskin fibroblast conditioned medium, wherein foreskin cells for said human foreskin fibroblast conditioned medium are not genetically modified, and wherein said human foreskin fibroblast conditioned medium being capable of maintaining said embryonic stem cells in an undifferentiated, proliferative state through at least 87 passages.

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

3. The method of claim 1, wherein said foreskin cells for said human foreskin fibroblast conditioned medium are obtained from an 8-14 day old male individual.

4. The method of claim 1, wherein said culturing is effected under culturing conditions including a growth medium which comprises serum and/or serum replacement.

5. The method of claim 4, wherein said serum and/or said serum replacement are provided at a concentration of at least 10%.

6. The method of claim 4, wherein said serum is human serum.

7. The method of claim 4, wherein said serum and/or said serum replacement are provided at a concentration of 15%.

8. The method of claim 1, wherein said culturing is effected under culturing conditions including a growth medium which comprises a factor selected from the group consisting of a growth factor, an anti oxidant and an amino acid.