Generation of neural stem cells from human trophoblast stem cells
Provided herein are isolated neural stem cells. Also provided are methods for treatment of neurodegenerative diseases using suitable preparations comprising the isolated neural stem cells.
1. A method of inducing a mammalian trophoblast stem cell to differentiate into a population of neural stem cells with reduced immunogenicity, comprising
contacting said mammalian trophoblast stem cell with an induction drug from about 3 hours to about 120 hours before removing said induction drug, wherein said induction drug comprises retinoic acid, and whereby said mammalian trophoblast stem cell is induced to differentiate into said population of neural stem cells with reduced immunogenicity such that said population of neural stem cells uniformly has an absence of CD33 protein expression and a low level of CD133 protein expression compared to a mammalian trophoblast stem cell not contacted with said induction drug.
2. The method of claim 1 , wherein said mammalian trophoblast stem cell is a human trophoblast stem cell.
3. The method of claim 1 , wherein said population of neural stem cells are human and, when administered to a human subject, does not induce an immune response or inhibits an immune response in said human subject.
4. The method of claim 1 , wherein said induction drug further comprises nicotinamide, beta-mercaptoethanol, vitamin B12, heparin, putrescine, biotin, Fe 2+ , butylated hydroxyanisole, valproic acid, forskolin, 5-azacytidine, indomethacin, isobutylmethylxanthine, or insulin.
5. The method of claim 1 , wherein said population of neural stem cells produces dopamine, a subunit of a glutamate NMDA receptor, synapsin I, a-calcium channel marker, GAP-43, voltage-dependent K + channel, a voltage-dependent Ca 2+ channel, or a voltage-dependent Na + channel.
6. The method of claim 1 , wherein said population of neural stem cells are human.
7. The method of claim 1 , wherein said induction drug is retinoic acid.
8. The method of claim 1 , wherein said population of neural stem cells does not induce an immune response or inhibits an immune response by a T cell, a B cell, a macrophage, a microglia cell, a mast cell, or a natural killer (NK) cell.
9. The method of claim 1 , wherein said induction drug decreases activity of Phosphatidylinositol 4,5-bisphosphate (PIP2) in the mammalian trophoblast stem cell.
10. The method of claim 1 , wherein said induction drug decreases expression of PIP2 in the mammalian trophoblast stem cell.
11. The method of claim 1 , wherein said induction drug decreases an amount of mRNA encoding PIP2 or an amount of a PIP2 protein translated from an mRNA in the mammalian trophoblast stem cell.
12. The method of claim 1 , wherein said induction drug activates Nuclear Factor of Activated T-cells (NFAT1) in the mammalian trophoblast stem cell.
13. The method of claim 12 , wherein said NFAT1 modulates microtubule assembly.
14. The method of claim 1 , wherein said contacting lasts from about 3 hours to about 24 hours.
15. The method of claim 1 , wherein said contacting lasts from about 3 hours to about 18 hours.
16. The method of claim 1 , further comprising removing said induction drug.
17. The method of claim 1 , further comprising washing said population of neural stem cells.
18. The method of claim 1 , further comprising washing said population of neural stem cells using phosphate buffer saline (PBS).