Primate embryonic stem cells

• US 20060040383A1
• Filed: 05/06/2003
• Published: 02/23/2006
• Est. Priority Date: 01/20/1995
• Status: Abandoned Application

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Thomson, James A.
• Application Number:
  US10/430,496
• Publication Number:
  US 20060040383A1
• Time in Patent Office: Days
• US Class Current: 435/363
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• 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

A purified preparation of primate embryonic stem cells is disclosed. This preparation is characterized by the following cell surface markers: SSEA-1 (−); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); and alkaline phosphatase (+). In a particularly advantageous embodiment, the cells of the preparation are human embryonic stem cells, have normal karyotypes, and continue to proliferate in an undifferentiated state after continuous culture for eleven months. The embryonic stem cell lines also retain the ability, throughout the culture, to form trophoblast and to differentiate into all tissues derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). A method for isolating a primate embryonic stem cell line is also disclosed.

Claims

1-11. -11. (canceled)

12. A method of making differentiated human cells comprising the steps of

(a) obtaining a culture of human embryonic stem cells, the embryonic stem cells having the ability to develop into mesoderm, endoderm and ectoderm;

and

(b) culturing the embryonic stem cells so that they differentiate into human cells selected from the group consisting of mesoderm, endoderm and ectoderm cells.

13. A method as claimed in claim 12 wherein the human embryonic stem cells are genetically engineered.

14. Human differentiated cells in culture made by the method of claim 12.

15. A method of making differentiated human cells in vitro comprising the steps of

(a) providing a culture of human embryonic stem cells, the embryonic stem cells having the properties of being euploid, maintenance of the potential to differentiate into mesoderm, endoderm and ectoderm, and that they are inhibited from differentiation when cultured on a fibroblast feeder layer;

and

(b) culturing the embryonic stem cells in the absence of a fibroblast feeder layer such that the cells differentiate into human cells selected from the group consisting of mesoderm, endoderm and ectoderm.

16. A method as claimed in claim 15 wherein the human cells are genetically engineered.