Derivation And Culture Of Human Embryo-Derived Cells

• US 20080050813A1
• Filed: 11/16/2005
• Published: 02/28/2008
• Est. Priority Date: 11/16/2004
• Status: Active Grant

• Current Assignee: Sonic Master Limited
• Original Assignee: Sydney Ivf Limited
• Inventors: Peura, Teija, Jansen, Robert Paul
• Granted Patent: US 9,359,590 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: C12N 5/0604 Whole embryos; Culture medium therefor
• 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 concerns methods for deriving and culturing embryonic cells and in particular to methods for maintaining the undifferentiated state of stems cells and cell lines in culture. The invention also concerns cells and cell lines derived by the methods of the invention.

Claims

1. Method of maintaining the undifferentiated state of an embryonic cell or embryonic cell line in culture comprising the step of culturing the embryo-derived cells or embryo-derived cell lines under conditions of low oxygen tension.

4. Method according to any one of claims 1 to 3, wherein the cell or cell line is an embryonic stem cell or embryonic stem cell line.

5. Method according to claim 4, wherein the embryonic stem cell or embryonic stem cell line is a bona fide human embryonic stem cell or embryonic stem cell line.

6. Method according to any one of claims 1 to 5, wherein the embryonic stem cell or embryonic stem cell line is derived from a human blastocyst.

10. Embryonic cell or embryonic cell line obtained according to the method of any one of claims 1 to 9.

11. Use of a cell or cell line according to claim 10, or a cell or cell line derived by a method of any one of claims 1 to 9, for pre-implantation genetic testing, or diagnosis of a genetic defect or chromosomal abnormality.

12. Use of a cell or cell line according to claim 10, or a cell or cell line derived by a method of any one of claims 1 to 9 for chromosomal karyotyping.

2. Method of supporting the proliferation of undifferentiated embryonic cell or embryonic cell line in culture comprising the step of culturing the embryo-derived cells or embryo-derived cell lines under conditions of low oxygen tension.

3. Method for large scale production of undifferentiated embryonic cells comprising the step of culturing embryo-derived cells or embryo-derived cell lines under conditions of low oxygen tension.

7. Method of derivation of embryo-derived cells or embryo-derived cell lines comprising the steps of:

a) culturing an embryo under low oxygen tension conditions for a time sufficient to produce embryonic cell-like outgrowths, and

b) isolating and culturing the embryonic cell-like outgrowths under low oxygen tension conditions to obtain embryo-derived cells or embryo-derived cell lines.

9. A method according to claim 7 or 8, wherein the embryo-derived cell or embryo-derived cell line is a stem cell or stem cell line

8. Method of derivation of embryo-derived cells or embryo-derived cell lines comprising the steps of:

a) incubating a putative zygote in a culture medium under low oxygen tension conditions to derive an embryo,

b) culturing embryo obtained in stop a) under low oxygen tension conditions for a time sufficient to produce embryonic cell-like outgrowths, and

c) isolating and culturing the embryonic cell-like outgrowths under low oxygen tension conditions to obtain embryo-derived cells or embryo-derived cell lines.