Mesenchymal cells derived from human embryonic stem cells

• US 20030036194A1
• Filed: 07/03/2002
• Published: 02/20/2003
• Est. Priority Date: 07/06/2001
• Status: Abandoned Application

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Inventors: Thies, R. Scott, Xu, Chunhui
• Application Number:
  US10/189,276
• Publication Number:
  US 20030036194A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/04 from germ cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12N 5/0654 Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0662 Stem cells

Abstract

This invention provides populations of mesenchymal cells obtained from pluripotent stem cells by differentiating them ex vivo. Multipotent mesenchymal cells can in turn be differentiated into more specialized cell types such as osteoblasts, with properties that make them suitable for reconstituting musculoskeletal cell function in an individual. The compositions, methods, and techniques described in this disclosure can be used for a variety of commercially important diagnostic, drug screening, and therapeutic applications.

Claims

1. A set of two isolated cell populations for generating human mesodermal cell populations, consisting of:

a first cell population comprising pluripotent stem (pPS) cells isolated from a human blastocyst, and

a second cell population that proliferates in culture, comprising at least ˜

30% pPS derived mesenchymal cells, identifiable by the criteria that they express either CD29 or CD44.

2. The set of cell populations of claim 1, wherein the mesenchymal cells express at least 3 of the following:

CD166 (ALCAM), CD29, CD44, GATA-4, and STRO-1.

3. The set of cell populations of claim 1, wherein the mesenchymal cells express either osteonectin or osteocalcin, and either type 1 collagen or vimentin.

4. The set of cell populations of claim 1, wherein the mesenchymal cells have increased alkaline phosphatase activity when cultured with bone morphogenic protein.

5. The set of cell populations of claim 1, wherein medium preconditioned by the mesenchymal cells causes proliferation of human embryonic stem cells without differentiation.

6. The set of cell populations of claim 1, wherein the osteoblasts or osteoblast precursors form an extracellular matrix comprising calcium when cultured in vitro.

7. The set of cell populations of claim 1, wherein the second cell population has been obtained by culturing the pPS cells so as to form embryoid bodies, plating the embryoid bodies onto a solid substrate, and then selecting cells having said criteria.

8. The set of cell populations of claim 1, wherein the second cell population has been genetically altered to express telomerase reverse transcriptase.

9. The set of cell populations of claim 1, wherein the pPS cells are a line of human embryonic stem cells.

11. A method of screening a compound for mesenchymal cell toxicity or modulation, comprising combining the compound with the second cell population of claim 1, and determining any mesenchymal cell toxicity or modulation resulting from the compound.

12. The method of claim 11, comprising determining whether the compound is toxic to the cell population.

13. The method of claim 11, comprising determining whether the compound affects proliferation of the cell population.

14. The method of claim 11, comprising determining whether the compound causes the second cell population to form progeny comprising at least ˜

10% osteoblasts, chondrocytes, hematopoietic cells, myoblasts, or adipocytes.

10. The human cell line designated HEF1.