In vitro and in vivo proliferation and use of multipotent neural stem cells and their progeny
First Claim
1. A method for the in vitro proliferation of a multipotent neural stem cell comprising the steps of:
- (a) obtaining neural tissue from a mammal, said neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia;
(b) dissociating said neural tissue to obtain a cell suspension comprising said multipotent neural stem cell;
(c) culturing said cell suspension in a first culture medium containing at least one proliferation-inducing growth factor to proliferate said neural stem cell and produce neural stem cell progeny; and
(d) passaging said neural stem cell progeny to a second culture medium containing at least one proliferation-inducing growth factor to further proliferate said neural stem cell progeny.
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Accused Products
Abstract
Nucleic acids may be obtained from neural cell cultures produced by using growth factors to induce the proliferation of multipotent neural stem cells. The resultant progeny may be passaged repeatedly to produce a sufficient number of cells to obtain representative nucleic acid samples. Clonal cultures may be produced. Nucleic acids may be obtained from both cultured normal and dysfunctional neural cells and from neural cell cultures at various stages of development. This information allows for the identification of the sequence of gene expression during neural development and can be used to reveal the effects of biological agents on gene expression in neural cells. Additionally, nucleic acids derived from dysfunctional tissue can be compared with that of normal tissue to identify genetic material which may be the cause of the dysfunction. This information could then be used in the design of therapies to treat the neurological disorder. A further use of the technology would be in the diagnosis of genetic disorders or for use in identifying neural cells at a particular stage in development.
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Citations
31 Claims
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1. A method for the in vitro proliferation of a multipotent neural stem cell comprising the steps of:
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(a) obtaining neural tissue from a mammal, said neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia;
(b) dissociating said neural tissue to obtain a cell suspension comprising said multipotent neural stem cell;
(c) culturing said cell suspension in a first culture medium containing at least one proliferation-inducing growth factor to proliferate said neural stem cell and produce neural stem cell progeny; and
(d) passaging said neural stem cell progeny to a second culture medium containing at least one proliferation-inducing growth factor to further proliferate said neural stem cell progeny. - View Dependent Claims (2, 3, 4, 5, 6)
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- 7. A cell culture comprising daughter multipotent neural stem cells that are the progeny of at least one parent multipotent neural stem cell obtained from dissociated mammalian neural tissue and proliferated in vitro in a culture medium containing at least one proliferation-inducing growth factor, wherein said daughter multipotent neural stem cells are capable of producing progeny that are capable of differentiating into neurons, and glia.
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9. A method of producing a cell culture comprising non-tumorigenic, genetically modified neural cells comprising the steps of:
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(a) dissociating neural tissue containing at least one multipotent neural stem capable of producing neural stem cell progeny that are capable of differentiating into neurons and glia, (b) proliferating said multipotent neural stem cell in a culture medium containing a proliferation-inducing growth factor to produce said neural stem cell progeny, and (c) genetically modifying said neural stem cell progeny to produce a cell culture comprising non-tumorigenic, genetically modified neural stem cell progeny. - View Dependent Claims (10, 11, 12, 13)
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14. A method of producing non-tumorigenic, genetically modified differentiated neural cells comprising the steps of:
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(a) isolating at least one multipotent neural stem cell from donor CNS tissue, said neural stem cell being capable of producing progeny that are capable of differentiating into neurons and glia, (b) proliferating the isolated neural stem cells in a culture medium containing a proliferation-inducing growth factor to produce precursor cells, (c) differentiating the precursor cells, and (d) genetically modifying said differentiated cells.
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15. A cell culture comprising non-tumorigenic, neural stem cell progeny that are capable of differentiating into neurons, and glia.
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16. A method of remyelinating a neuron comprising the steps of:
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(a) dissociating mammalian neural tissue containing at least one multipotent stem cell capable of producing neural stem cell progeny that are capable of differentiating into neurons, and glia, (b) exposing said multipotent stem cell to a culture medium containing at least one proliferation-inducing growth factor to produce neural stem cell progeny, (c) harvesting said neural stem cell progeny, and (d) causing said harvested neural stem cell progeny to come into contact with a demyelinated axon to effect remyelination of said demyelinated axon. - View Dependent Claims (17)
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18. A method of remyelinating a neuron comprising the steps of:
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(a) dissociating mammalian neural tissue containing at least one multipotent stem cell capable of producing neural stem cell progeny that are capable of differentiating into neurons, astrocytes, and oligodendrocytes, (b) exposing said multipotent stem cell to a first culture medium containing at least one proliferation-inducing growth factor to produce neural stem cell progeny, (c) differentiating said neural stem cell progeny in a second culture medium that is substantially free of said proliferation-inducing factor to produce oligodendrocytes, and (d) causing said oligodendrocytes to come into contact with a demyelinated axon to effect remyelination of said demyelinated axon.
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19. A method for the in vivo proliferation of a precursor cell located in the CNS tissue of a mammal, said method comprising administering at least one proliferation-inducing growth factor to said CNS tissue to induce the proliferation of said cell.
- 20. A method for the in vivo genetic modification of a CNS precursor cell located in tissue lining a CNS ventricle of a mammal, said method comprising administering genetic material to said CNS ventricle to infect said cells, said genetic material being capable of encoding at least one neurological agent.
- 22. A method of treating a neurological disorder of a mammal comprising administering a composition comprising a proliferation-inducing growth factor to a ventricle of the central nervous system of said mammal, said ventricle being lined by tissue comprising at least one precursor cell, said growth factor inducing the in vivo proliferation and, optionally, differentiation of said precursor cell to form tissue comprising proliferated precursor cells.
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25. A method of treating a neurological disorder of a mammal comprising administering genetic material to a CNS ventricle of a mammal, said ventricle being lined by tissue comprising at least one precursor cell, said precursor cell being genetically modified in vivo by said genetic material, said genetic material being capable of encoding at least one neurological agent.
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26. A method of transplanting neural stem cell progeny to a host comprising:
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(a) obtaining neural tissue from a mammal, said neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons, and glia;
(b) dissociating said neural tissue to obtain a cell suspension comprising said multipotent neural stem cell;
(c) culturing said cell suspension in a culture medium containing at least one proliferation-inducing growth factor to proliferate said neural stem cell and produce neural stem cell progeny; and
(d) transplanting said neural stem cell progeny to said host. - View Dependent Claims (27)
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28. A method for determining the effect of at least one biological agent on neural precursor cells comprising:
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(a) dissociating mammalian neural tissue containing at least one multipotent stem cell, (b) proliferating said multipotent stem cell in a culture medium containing at least one growth factor to obtain a culture of proliferated precursor cells, (c) contacting said proliferated precursor cells with said biological agent, and (d) determining the effects of said biological agent on said precursor cells.
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29. A method for determining the effect of at least one biological agent on the differentiation of neural cells comprising:
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(a) dissociating mammalian neural tissue containing at least one multipotent stem cell, (b) proliferating said multipotent stem cell in a first culture medium containing at least one growth factor to obtain a culture of proliferated precursor cells, (c) inducing said proliferated precursor cells to differentiate in a second culture medium in the presence said biological agent, and (d) determining the effects of said biological agent on the differentiation of said precursor cells.
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30. A method for determining the effect of at least one biological agent on differentiated neural cells comprising:
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(a) dissociating mammalian neural tissue containing at least one multipotent stem cell, (b) proliferating said multipotent stem cell in a first culture medium containing at least one growth factor to obtain a culture of proliferated precursor cells, (c) inducing said proliferated precursor cells to differentiate into differentiated neural cells, (d) contacting said differentiated neural cells with said biological agent, and (d) determining the effects of said biological agent on said differentiated neural cells.
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31. A cDNA library prepared from neural stem cell progeny.
Specification