Oligodendrocytes derived from human embryonic stem cells for remyelination and treatment of spinal cord injury

• US 7,285,415 B2
• Filed: 04/04/2003
• Issued: 10/23/2007
• Est. Priority Date: 07/11/2002
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Keirstead, Hans S., Nistor, Gabriel I.
• Primary Examiner(s): Hayes; Robert C.
• Assistant Examiner(s): KOLKER, DANIEL E
• Application Number:
  US10/406,817
• Publication Number:
  US 20040009593A1
• Time in Patent Office: 1,663 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/395 Thyroid hormones
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0622 Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

This invention provides populations of neural cells bearing markers of glial cells, such as oligodendrocytes and their precursors. The populations are generated by differentiating pluripotent stem cells such as human embryonic stem cells under conditions that promote enrichment of cells with the desired phenotype or functional capability. Various combinations of differentiation factors and mitogens can be used to produce cell populations that are over 95% homogeneous in morphological appearance, and the expression of oligodendrocyte markers such as GalC. The cells are capable of forming myelin sheaths, and can be used therapeutically improve function of the central nervous system.

Claims

1. A method for producing glial cells from human embryonic stem (hES) cells, comprising culturing hES cells in medium comprising a growth factor, a ligand for a thyroid hormone receptor, and a ligand for a retinoic acid receptor.

2. The method of claim 1, wherein the growth factor is basic fibroblast growth factor.

3. The method of claim 1, wherein the hES cells are cultured in a medium comprising thyroid hormone T3 and retinoic acid.

4. The method of claim 1, wherein the hES cells are cultured in a medium comprising the thyroid hormone T3 and selenium.

5. The method of claim 1, wherein the hES cells are cultured in suspension so as to form yellow spheres.

6. The method of claim 1, further comprising separating glial cells from non-glial cells.

7. The method of claim 6, wherein the separating is performed by plating cells from the culture onto a solid surface, and harvesting cells that adhere to the surface.

8. The method of claim 1, further comprising harvesting from the culture a cell population in which at least 80% of cells are positive for NG2 proteoglycan, but negative for NeuN.

9. The method of claim 1, further comprising culturing the glial cells in the absence of mitogens so as to produce a population containing cells that have numerous processes characteristic of mature oligodendrocytes.

10. The method of claim 1, further comprising formulating the glial cells in a composition suitable for administration into a spinal cord.

11. The method of claim 1, further comprising screening the effect of a compound by combining the glial cells with the compound, and determining any resulting phenotypic or functional change in the cells.