Embryonic stem cells and neural progenitor cells derived therefrom
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Abstract
The present invention provides undifferentiated human embryonic stem cells, methods of cultivation and propagation and production of differentiated cells. In particular it relates to the production of human ES cells capable of yielding somatic differentiated cells in vitro, and committed progenitor cells such as neural progenitor cells capable of giving rise to mature somatic cells including neural cells and/or glial cells and uses thereof. The invention also provides methods that generate in vitro and in vivo models of controlled differentiation of ES cells towards the neural lineage. The model and the cells generated along the pathway of neural differentiation may be used for the study of the cellular and molecular biology of human neural development, for the discovery of genes, growth factors, and differentiation factors that play a role in neural differentiation and regeneration, for drug discovery and for the development of screening assays for teratogenic, toxic and neuroprotective effects.
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Citations
65 Claims
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1-7. -7. (canceled)
- 8. A differentiated committed human progenitor cell line capable of differentiation and propagation into mature neurons or glial cells said cell line derived from undifferentiated human embryonic stem cells.
- 11. A neural progenitor cell differentiated in vitro from an undifferentiated human embryonic stem cell.
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28-38. -38. (canceled)
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39. A method of inducing somatic differentiation of stem cells in vitro into progenitor cells said method comprising:
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obtaining undifferentiated embryonic stem cells; and
providing a differentiating signal under conditions which are non-permissive for stem cell renewal, do not kill cells and/or induces unidirectional differentiation toward extraembryonic lineages. - View Dependent Claims (46, 47, 48, 49, 52)
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40-45. -45. (canceled)
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50. A method of inducing somatic cells from embryonic stem cell derived somatic progenitors, said method comprising:
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obtaining a source of embryonic stem cell derived somatic progenitors;
culturing the progenitor cells on an adhesive substrate; and
inducing the cells to differentiate to somatic cells under conditions which favour somatic differentiation. - View Dependent Claims (53, 54, 55)
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51. (canceled)
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56-63. -63. (canceled)
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64. A method of producing an enriched preparation of human ES derived neural progenitor cells, said method comprising:
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obtaining an undifferentiated human embryonic stem cell comprising obtaining an in vitro fertilised human embryo and growing the embryo to a blastocyst stage of development;
removing inner cells mass (ICM) cells from the embryo;
culturing ICM cells under conditions which do not induce extraembryonic differentiation and cell death, and promote proliferation of undifferentiated stem cells;
recovering stem cells;
inducing somatic differentiation of the embryonic stem cell to a neural progenitor cell comprising obtaining undifferentiated embryonic stem cells;
providing a differentiating signal under conditions which are non-permissive for stem cell renewal, do not kill cells or induces unidirectional differentiation toward extraembryonic lineages;
identifying a neural progenitor cell by expressed markers of primitive neuroectoderm and neural stem cells such as polysialyated N-CAM, intermediate filament proteins such as nestin and vimentin and the transcription factor Pax-6; and
culturing the neural progenitor cells to promote proliferation and propagation.
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65-85. -85. (canceled)
Specification