Method and Medium for Neural Differentiation of Pluripotent Cells

• US 20110229441A1
• Filed: 12/07/2009
• Published: 09/22/2011
• Est. Priority Date: 12/05/2008
• Status: Abandoned Application

• Current Assignee: Association Francaise Contre Les Myopathies, Inserm (institut national de la sant et de la recherche mdicale
• Original Assignee: Association Francaise Contre Les Myopathies
• Inventors: Benchoua, Alexandra, Aubry, Laetitia, Perrier, Anselme
• Application Number:
  US12/994,980
• Publication Number:
  US 20110229441A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61K 35/30 Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
• 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: C12N 2500/05 Inorganic components
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/40 Nucleotides, nucleosides or bases
• 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 2501/16 Activin; Inhibin; Mullerian inhibiting substance
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0619 Neurons
• CPC Class Codes: C12N 5/0623 Stem cells
• CPC Class Codes: G01N 33/5058 Neurological cells

Abstract

The invention relates to a culture medium comprising an inhibitor of the BMP signaling pathway; and an inhibitor of the TGF/activin/nodal signaling pathway and to a method for obtaining a population of neural precursors using said culture medium.

Claims

1. A culture medium comprising:

a) an inhibitor of the Bone Morphogenetic Protein (BMP) signaling pathway; and

b) an inhibitor of the Transforming Growth Factor (TGF)/activin/nodal signaling pathway.

2. A culture medium according to claim 1, wherein said culture medium comprises:

a) a source of carbon;

b) essential amino-acids;

c) vitamins;

d) a purine and a pyrimidine;

e) inorganic salts;

f) a molecule known to limit natural ageing;

g) an antioxidant;

h) a phospholipid precursor;

i) an unique fatty acid; and

j) a carrier protein.

3. A culture medium according to claim 1, wherein said inhibitor of the BMP signaling pathway is selected from the group consisting of noggin, chordin and follistatin and variants and fragments thereof which inhibit the BMP signaling pathway.

4. A culture medium according to claim 1, wherein said inhibitor of the TGF/activin/nodal signaling pathway is selected from the group consisting of SB431542, Lefty-A and Cerberus and variants and fragments of Lefty-A and Cerberus which inhibit the TGF/activin/nodal signaling pathway.

5. A method for producing a population of neural precursors wherein said method comprises the step of culturing pluripotent cells with the culture medium as defined in claim 1.

6. A method according to claim 5 wherein said pluripotent cells are human pluripotent cells.

7. A method according to claim 5 wherein said pluripotent cells are stem cells.

8. A method according to claim 7 wherein said stem cells are embryonic stem cells.

9. A method according to claim 5 wherein said pluripotent cells are induced pluripotent cells (IPS).

10. A population of neural precursors obtained by a method as defined in claim 5.

11. Method for obtaining a population of neurons wherein said method comprises the steps of:

a) producing a population of neural precursors according to the method of claim 5;

b) differentiating said population of neural precursors into neurons.

12. A population of neurons obtained by the method of claim 11.

14. A method for screening compounds having a neuroprotective and/or neurotoxic effect wherein said method comprises the steps of:

a) Culturing a population of neural precursors according to claim 10 in the presence of a test compound;

b) Comparing the survival of the cells of step a) to that of a population of neural precursors as defined above cultured in the absence of said test compound.

15. A method for screening compounds having a neuroprotective and/or neurotoxic effect wherein said method comprises the steps of:

a) culturing a population of neurons according to claim 12 in the presence of a test compound;

b) comparing the survival of the cells of step a) to that of a population of neurons as defined above cultured in the absence of said test compound.

16. A method for treating a neurodegenerative disease or brain injury comprising the step of administering a pharmaceutically effective amount of a population of neural precursors according to claim 10 to a patient in need thereof.

17. A method for treating a neurodegenerative disease or brain injury comprising the step of administering a pharmaceutically effective amount of a population of neurons according to claim 12 to a patient in need thereof.

13. (canceled)