Transplantation of human neural cells for treatment of neurodegenerative conditions
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.
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 pg/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 8 , wherein the enzymatic dissociation includes treating the cells with trypsin.
10. The method of claim 1 , wherein a therapeutically effective amount of said expanded cell population is introduced to at least one area of a recipient spinal cord to treat a neurodegenerative condition.
11. The method of claim 10 , wherein the neurodegenerative condition is amyotrophic lateral sclerosis (ALS).