In vitro growth and proliferation of multipotent neural stem cells and their progeny
AssigneeNeurospheres, Ltd.
InventorsSamuel Weiss, Brent Reynolds, Joseph P. Hammang, E. Edward Baetge
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Abstract
A method for the in vitro proliferation and differentiation of neural stem cells and stem cell progeny comprising the steps of (a) isolating the cells from a mammal, (b) exposing the cells to a culture medium containing a growth factor, (c) inducing the cells to proliferate, and (d) inducing the cells to differentiate is provided.
Claims80
A method for preparing a population of mammalian neural cells enriched with multipotent neural stem cells comprising:
- (a) obtaining a population of mammalian neural cells which contains at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes;
- (b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation;
- and
- (c) combining the population of mammalian neural cells obtained in (a) with the culture medium prepared in (b) and culturing said mammalian neural cells under conditions that allow the proliferation of said at least one multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells to produce a cell culture that contains a percentage of multipotent neural stem cells that is at least ten fold higher than that of said population of mammalian neural cells obtained in (a).
The method of claim additionally comprising:
- (d) preparing at least one subsequent cell culture by combining said multipotent neural stem cell progeny produced in (c) with fresh culture medium containing one or more predetermined growth factors which induces multipotent neural stem cell proliferation to proliferate said daughter multipotent neural stem cells to produce a non-primary cell culture that contains a percentage of multipotent neural stem cells that is at least ten fold higher than that of said cell culture prepared in (c).
A method for inducing the in vitro proliferation of a multipotent neural stem cell comprising:
- (a) obtaining cells derived from juvenile or adult mammalian neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia;
- (b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation;
- and
- (c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells.
The method of 10 wherein step (d) is repeated.
A method for inducing the in vitro proliferation of a multipotent neural stem cell comprising:
- (a) obtaining cells derived from mammalian neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes, wherein said obtained cells have not been cultured in a serum-containing medium;
- (b) preparing a substantially serum-free culture medium containing at one or more predetermined growth factors capable of inducing proliferation of said multipotent neural stem cell;
- (c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells;
- and
- (d) preparing at least one subsequent cell culture by combining said multipotent neural stem cell progeny with fresh substantially serum-free culture medium containing at least one or more predetermined growth factors which induces multipotent neural stem cell proliferation to proliferate said daughter multipotent neural stem cells to produce more progeny which include more daughter multipotent neural stem cells.
A method for the in vitro proliferation of a human multipotent neural stem cell comprising:
- (a) obtaining cells derived from human neural tissue containing at least one multipotent neural stem cell capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes;
- (b) preparing a culture medium containing one or more predetermined growth factors capable of inducing multipotent neural stem cell proliferation;
- and
- (c) preparing a cell culture by combining the cells obtained in (a) with the culture medium prepared in (b) to induce proliferation of said multipotent neural stem cell to produce multipotent neural stem cell progeny which includes daughter multipotent neural stem cells.
A composition comprising a population of non-primary neural cells which are derived from a primary cell culture, said population of non-primary. neural cells having a greater percentage of multipotent neural stem cells compared to that of said primary cell culture, wherein a single multipotent neural stem cell is capable of producing progeny that are capable of differentiating into neurons and glia, including astrocytes.
A cluster of cells prepared in vitro consisting of the progeny of a single multipotent neural stem cell, wherein said progeny are capable of differentiating into neurons and glia, including astrocytes.
Dependencies are inferred from the claim text.