Pluripotent cells that are immunopositive for both the neural progenitor marker nestin and a pluripotent cell marker are provided. The cells exhibit rapid doubling times and can be maintained in vitro for extended periods. Also provided are cell cultures containing the pluripotent cells, a method of
Pluripotent cells that are immunopositive for both the neural progenitor marker nestin and a pluripotent cell marker are provided. The cells exhibit rapid doubling times and can be maintained in vitro for extended periods. Also provided are cell cultures containing the pluripotent cells, a method of transplanting human pluripotent cells to a host, and a method of reducing seizure activity in a subject. These pluripotent cells, when transplanted into the ventricle of a host animal, migrate to the site of damage and adopt a suitably corrective phenotype, resulting in both structural and functional restoration.
대표청구항▼
1. A method of transplanting progenitor cells to a mammalian host, comprising: (a) obtaining a population of isolated mammalian progenitor cells that co-express nestin and Oct-4 from fetal telencephalon and/or mesencephalon, and a culture medium that has a total calcium concentration of 0.03 to 0.15
1. A method of transplanting progenitor cells to a mammalian host, comprising: (a) obtaining a population of isolated mammalian progenitor cells that co-express nestin and Oct-4 from fetal telencephalon and/or mesencephalon, and a culture medium that has a total calcium concentration of 0.03 to 0.15 mM, wherein the progenitor cells are suspended in the culture medium and wherein the progenitor cells continue to proliferate in an undifferentiated state for at least 4 months; and(b) transplanting the progenitor cells to the mammalian host, wherein said progenitor cells further express a marker selected from the group consisting of TRA-1-60, TRA-1-81 and SSEA-4. 2. The method of claim 1, wherein the isolated mammalian cells express each of TRA-1-60, TRA-1-81 and SSEA-4. 3. The method of claim 1, wherein the cells have a doubling rate of less than 12 days. 4. The method of claim 1, wherein the cells have a doubling rate of about 5 days. 5. The method of claim 1, wherein the cells continue to proliferate for 2 years in vitro. 6. The method of claim 1, wherein the subject is human, equine, canine, feline, porcine, ovine or rodent. 7. The method of claim 1, wherein the medium has a total calcium concentration of less than 0.1 mM. 8. The method of claim 7, wherein the total calcium concentration is about 0.05 mM. 9. The method of claim 1, wherein the medium further comprises: (a) about 15-100 ng/μl epidermal growth factor (EGF);(b) about 10-150 ng/μl basic fibroblast growth factor (bFGF);(c) about 10-75 ng/μl transforming growth factor-alpha (TGFα). 10. The method of claim 9, wherein the medium further comprises: (d) about 25-150 ng/μl leukemia inhibiting factor (LIF). 11. The method of claim 10, wherein the LIF is about 25 ng/μl. 12. The method of claim 9, wherein the EGF is about 20 ng/μl. 13. The method of claim 9, wherein the bFGF is about 10 ng/μl. 14. The method of claim 9, wherein the TGFα is about 10 ng/μl. 15. The method of claim 9, wherein the culture medium is free of a feeder layer. 16. The method of claim 9, wherein the culture medium is serum-free. 17. The method of claim 9, further comprising 0.5-2.5% B27 supplement. 18. The method of claim 9, wherein the growth factors EGF, bFGF and TGFα are recombinant growth factors. 19. The method of claim 9, wherein the cells and the growth factors are human. 20. The method of claim 9, wherein the culture medium further comprises about 0.11 mg/ml sodium pyruvate.
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