Transplantation of human neural cells for treatment of neurodegenerative conditions

• US 7,691,629 B2
• Filed: 11/17/2005
• Issued: 04/06/2010
• 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: Hazel, Thomas G., Johe, Karl K.
• Primary Examiner(s): Ketter; James S
• Application Number:
  US11/281,640
• Publication Number:
  US 20060141622A1
• Time in Patent Office: 1,601 Days
• Field of Search: None
• US Class Current: 435/402
• 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 culturing human neural stem cells, said method comprising:

providing human neural stem cells;

incubating a culture vessel with an about 0.1 μ

g/mL to about 1 mg/mL solution of poly-D-lysine for about 5 minutes to about 3 hours;

washing and drying the culture vessel to form a pre-coated culture vessel;

seeding the neural stem cells in said pre-coated culture vessel in absence of serum;

adding a fibronectin solution and at least one mitogen to the pre-coated culture vessel and culturing the neural stem cells in absence of serum;

further culturing the neural stem cells with at least one mitogen; and

passaging the cultured neural stem cells prior to confluence.

2. The method of claim 1, wherein the mitogen includes bFGF.

3. The method of claim 1, wherein the mitogen includes EGF.

4. The method of claim 1, wherein the mitogen includes TGF-alpha.

5. The method of claim 1, wherein the mitogen includes aFGF.

6. The method of claim 1 wherein the solution of poly-D-lysine has a poly-D-lysine concentration of about 100 μ

g/mL.

7. The method of claim 1 wherein the step of incubating a culture vessel with a solution of poly-D-lysine is performed for about 1 hour.

8. The method of claim 1 wherein the fibronectin solution has a fibronectin concentration of about 1 μ

g/mL.

9. The method of claim 1 wherein the fibronectin solution and the at least one mitogen are added to the pre-coated culture vessel substantially at the time of seeding.

10. The method of claim 1 wherein the provided neural stem cells are derived from human embryonic stem cells.

11. The method of claim 1 wherein the provided neural stem cells are from a central nervous system tissue.

12. The method of claim 11 wherein the central nervous system tissue comprises spinal cord tissue.

13. The method of claim 12 wherein the spinal cord tissue is obtained from a post-mortem fetus having a gestational age of about 6.5 to about 20 weeks.