Transplantation of human neural cells for treatment of neurodegenerative conditions

• US 20060141622A1
• Filed: 11/17/2005
• Published: 06/29/2006
• Est. Priority Date: 11/17/2004
• Status: Active Grant

• Current Assignee: Neuralstem Incorporated (City of Tianjin (China))
• Original Assignee: Neuralstem Incorporated (City of Tianjin (China))
• Inventors: Johe, Karl K., Hazel, Thomas G.
• Granted Patent: US 7,691,629 B2
• Time in Patent Office: Days
• US Class Current: 435/368
• 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: A61K 9/0085 Brain, e.g. brain implants; Spinal cord
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• 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 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/113 Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/148 Transforming growth factor alpha [TGF-a]
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

A method of treating neurodegenerative conditions is provided. Neural stem cells may be implanted at and/or remote from a region of neuron degeneration. The methods can include isolating neural stem cells from regions where specific types of neurons corresponding to the neurons to be replaced are generated. The methods can include isolating neural stem cells secreting growth factors affecting the growth and/or regeneration of specific types of neuron. In this invention, we disclose a method of treating such disorders, including several neurodegenerative disorders arising from the lack of cells that produce particular neurotransmitters in neural circuitry by transplanting exogenously cultured and expanded neural progenitors which, upon transplantation into a neural tissue, differentiate into neurons capable of integrating and producing neurotransmitters in sufficient quantities and in a sufficient manner to overcome the symptoms associated with the neurodegeneration.

Claims

1. A method of expanding in vitro at least one neural stem cell to an expanded population of neural stem cells, wherein each neural stem cell expansion exceeds thirty cell doublings without differentiating, said method comprising:

dissociating neural stem cells from central nervous system tissue;

providing at least one extracellular protein to a culture vessel, wherein the extracellular protein includes at least about 10 μ

g/mL of poly-D-lysine and less than about 1 mg/ml fibronectin;

culturing the dissociated neural stem cells in said culture vessel in the absence of serum;

adding to the culture vessel at least one growth factor; and

passaging the cultured cells prior to confluence.

2. The method of claim 1, wherein the expanded neural stem cells are capable of differentiating into neurons.

3. The method of claim 1, wherein expanding said neural stem cell includes adding fibronectin to culture medium as a soluble factor.

4. The method of claim 1, wherein the growth factor includes bFGF.

5. The method of claim 1, wherein the growth factor includes EGF.

6. The method of claim 1, wherein the growth factor includes TGF-alpha.

7. The method of claim 1, wherein the growth factor includes aFGF.

8. The method of claim 1, wherein dissociating the cells and passaging the cells includes enzymatic dissociation.

9. The method of claim 4, wherein the enzymatic dissociation includes treating the cells with trypsin.

10. The method of claim 1, wherein a therapeutically effective amount of said expanded population is introduced to at least one area of a recipient nervous system to treat a neurodegenerative condition.