Methods and compositions relating to embryonic stem cell lines

• US 20060275899A1
• Filed: 12/22/2005
• Published: 12/07/2006
• Est. Priority Date: 12/30/2004
• Status: Abandoned Application

• Current Assignee: Stemlifeline, Inc.
• Original Assignee: Stemlifeline, Inc.
• Inventors: Krtolica, Ana, Foulk, Russell A., Simon Valles, Carlos
• Application Number:
  US11/317,886
• Publication Number:
  US 20060275899A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: C12N 2500/02 Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2502/243 Cells of the female genital tract, non-germinal ovarian cells
• 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 invention provides methods for generating customized hESC and lines thereof for future use in the treatment of genetic relations. In one aspect, the customized hESC lines are grown using human feeder cells or human serum from biological or genetic relations in order to reduce or preferably avoid contamination by foreign pathogens or exposure to foreign antigens.

Claims

1. A method for generating a human embryonic stem cell line comprising culturing stem cells from a human blastocyst generated using a female donor egg, on or in the presence of human feeder cells or human serum to generate a human embryonic stem cell line,

wherein the human feeder cells or human serum are derived from a biological or genetic relation of the blastocyst and wherein the human feeder cells are mitotically inactivated.

2. The method of claim 1, wherein the stem cells are generated by culturing inner cell mass (ICM) cells from the human blastocyst on or in the presence of the human feeder cells or human serum, growing stem cell-like colonies from the ICM cells on or in the presence of the human feeder cells or human serum, and isolating and culturing cells from the stem cell-like colonies on or in the presence of the human feeder cells or human serum.

3. The method of claim 1, wherein the human blastocyst is generated from a zygote using a female donor egg and a male donor sperm.

4. The method of claim 1, wherein the human blastocyst is generated by culturing a zygote in the presence of endometrial epithelial cells.

5. The method of claim 1, wherein the human feeder cells or human serum are derived from the female donor.

6. The method of claim 1, wherein the human feeder cells or human serum are derived from a biological non-genetic mother.

7. The method of claim 5, wherein the human feeder cells are amniotic epithelial cells, breast fibroblasts, endometrial stromal cells, fallopian tube fibroblasts, granulosa cells, oviduct fibroblasts or placental fibroblasts.

8. The method of claim 3, wherein the human feeder cells or human serum are derived from the male donor.

9. The method of claim 1, wherein the human feeder cells or human serum are derived from a sibling of the blastocyst.

10. The method of claim 5, wherein the human feeder cells are human bone marrow cells, human lung fibroblasts, human muscle cells, human oral fibroblasts, human skin fibroblasts, or tissue-derived stromal cells.

11. The method of claim 1, wherein the embryonic stem cell line is generated using human feeder cells.

12. The method of claim 1, wherein the embryonic stem cell line is generated using human serum.

13. The method of claim 1, wherein the embryonic stem cell line is generated using feeder-free culture conditions.

14. The method of claim 1, wherein the embryonic stem cell line is generated under a hypoxic condition.

15. The method of claim 14, wherein the hypoxic condition is 2%-5% oxygen content.

16. The method of claim 14, wherein the hypoxic condition is greater than 5%.

17. The method of claim 1, further comprising cryopreserving the human embryonic stem cell line, the human feeder cells, and/or the human serum.

18. The method of claim 17, wherein the human embryonic stem cell line, the human feeder cells and/or the human serum are cryopreserved separately.

19. A cell line generated by the method of claim 1.

20. A human embryonic stem cell line bank comprising a sample of a human embryonic stem cell line generated as in claim 1.

29. A method for determining therapeutic efficacy of a compound comprising

exposing a human embryonic stem cell line generated by the method of claim 1 with a compound, and

determining an effect of the compound on the embryonic stem cell line.

21. -28. (canceled)

30. -36. (canceled)