Hematopoietic differentiation of human pluripotent embryonic stem cells

• US 6,280,718 B1
• Filed: 11/08/1999
• Issued: 08/28/2001
• Est. Priority Date: 11/08/1999
• Status: Expired due to Term

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Thomson, James A., Kaufman, Dan S.
• Primary Examiner(s): CROUCH, DEBORAH
• Application Number:
  US09/435,578
• Time in Patent Office: 659 Days
• Field of Search: 424/93.1, 424/93.7, 435/325, 435/366, 435/375
• US Class Current: 424/93.1
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2502/1394 Bone marrow stromal cells; whole marrow
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

Disclosed herein are methods of obtaining human hematopoietic cells from human pluripotent embryonic stem cells using mammalian stromal cells. Hematopoietic cells derived in this way are useful for creating cell cultures suitable for transplantation, transfusion, and other purposes.

Claims

1. A method for obtaining human hematopoietic cells, comprising exposing a human pluripotent embryonic stem cell culture to mammalian hematopoietic stromal cells so as to thereby produce human hematopoietic cells, wherein at least some of the human hematopoietic cells that are so produced will form hematopoietic cell colony forming units if placed in methylcellulose culture.

2. The method of claim 1, wherein at least some of the human hematopoietic cells that are so produced are CD34⁺.

3. The method of claim 1, wherein the stromal cells are selected from the group consisting of bone marrow cells and embryonic yolk cells.

4. A method for obtaining human hematopoietic cells, comprising exposing a human pluripotent embryonic stem cell culture in vitro to mammalian hematopoietic stromal cells so as to thereby produce human hematopoietic cells, wherein at least some of the human hematopoietic cells that are so produced will form erythroid blast forming units if placed in methylcellulose culture.

5. The method of claim 4, wherein the human hematopoietic cells comprise at least some hematopoietic cells that are CD34⁺.

6. A method of transplanting human cells into a human recipient host, comprising:

obtaining human hematopoietic cells which have been derived in vitro from a human pluripotent embryonic stem cell culture;

obtaining a selected human cell other than hematopoietic cells, the selected non-hematopoietic cell having major histocompatibility complex compatibility to the human hematopoietic cells; and

transplanting both the human hematopoietic cells and selected human non-hematopoietic cell into the human host.

7. The method of claim 6, wherein the selected human cell is a human pancreatic islet.

8. The method of claim 6, wherein the selected human cell is a human oligodendroeytcs.

9. A method for obtaining human hematopoietic cells, comprising exposing a human pluripotent stem cell culture to mammalian hematopoietic stromal cells so as to thereby produce human hematopoietic cells, wherein at least some of the human hematopoietic cells that are so produced will form hematopoietic cell colony forming units if placed in methylcellulose culture.

10. A method of transplanting human cells into a human recipient host, comprising:

obtaining human hematopoietic cells which have been derived in vitro from a human pluripotent stem cell culture;

obtaining a selected human cell other than hematopoietic cells, the selected non-hematopoietic cell having major histocompatibility complex compatibility to the human hematopoietic cells; and

transplanting both the human hematopoietic cells and selected human non-hematopoietic cell into the human host.