Enriched central nervous system stem cell and progenitor cell populations, and methods for identifying, isolating and enriching for such populations
AssigneeStemCells California, Inc.
InventorsDavid W. Buck, Nobuko Uchida, Irving Weissman
Life of this patent
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Abstract
Enriched neural stem and progenitor cell populations, and methods for identifying, isolating and enriching for neural stem cells using reagent that bind to cell surface markers, are provided.
Claims48
A method for producing a population highly enriched for human central nervous system stem cells (CNS-SC) which can initiate neurospheres (NS-IC), comprising:
- (a) contacting a population containing neural or neural-derived cells with a reagent that recognizes a determinant on a cell surface marker recognized by monoclonal antibody AC133 or by monoclonal antibody 5E12;
- and
- (b) selecting for cells in which there is contact between the reagent and the determinant on the surface of the cells of step (a), to produce a population highly enriched for CNS-SC.
A method for enriching a population of neural cells for the population's neurosphere initiating stem cell (NS-IC) fraction, comprising:
- (a) combining a population comprising neural cells or neural-derived cells containing a fraction of NS-ICs with a reagent that recognizes a determinant on a cell surface marker recognized by monoclonal antibody AC133 or by monoclonal antibody 5E12;
- and
- (b) selecting for AC133+ and/or 5E12+ cells, wherein the selected AC133+ or 5E12+ cells are enriched in the fraction of NS-IC as compared with the population of neural cells.
A method for identifying a neurosphere initiating stem cell (NS-IC), comprising:
- (a) contacting a population containing neural or neural-derived cells with a reagent that recognizes a determinant on a cell surface marker recognized by monoclonal antibody AC133 or by monoclonal antibody 5E12;
- and
- (b) detecting the contact between a cell of the population containing neural or neural-derived cells and the reagent that recognizes a determinant on a cell surface marker recognized by monoclonal antibody AC133 or by monoclonal antibody 5E12, wherein an identification of a cell as being AC133+ or 5E12+ identifies the cell as an NS-IC.
A method for isolating a neurosphere initiating stem cell (NS-IC), comprising:
- (a) combining a population comprising neural cells or neural-derived cells containing a fraction of NS-ICs with a reagent that recognizes a determinant on a cell surface marker recognized by monoclonal antibody AC133 or by monoclonal antibody 5E12;
- (b) selecting AC133+ or 5E12+ cells, wherein the selected AC133+ or 5E12+ cells are enriched in the fraction of NS-ICs as compared with the population of neural cells;
- (c) introducing at least one AC133+ or 5E12+ cell to a culture medium capable of supporting the growth of NS-IC;
- and
- (d) proliferating the AC133+ or 5E12+ cell in the culture medium.
An in vitro cell culture composition comprising:
- (a) a population enriched in AC133+ CD45 cells or 5E12+ CD45− cells;
- and
- (b) a medium capable of supporting the growth the cells.
An in vitro cell culture composition comprising:
- (a) a population enriched in AC133+ CD45− CD34− cells or 5E12+ CD45− CD34− cells;
- and
- (b) a medium capable of supporting the growth the cells.
An in vitro cell culture composition comprising:
- (a) a population enriched in AC133+ 8G1−/lo cells or 5E12+ 8G1hi cells;
- and
- (b) a medium capable of supporting the growth the cells.
An in vitro cell culture composition comprising:
- (a) a population enriched in AC133+ 8G1hi cells or 5E12+ 8G1hi cells;
- and
- (b) a medium capable of supporting the growth the cells.
An in vitro cell culture composition comprising:
- (a) a population comprising at least 50% AC133− or 5E12+ neurosphere initiating cells (NS-IC) which stain positive for nestin and, in the presence of differentiation-inducing conditions, produce progeny cells that differentiate into neurons, astrocytes, and oligodendrocytes;
- and
- (b) a medium capable of supporting the growth of NS-IC.
A method for characterizing a neurosphere initiating stem cell (NS-IC), comprising:
- (a) introducing an isolated AC133+ or 5E12+ cell to a culture medium capable of supporting the growth of NS-IC;
- (b) proliferating the AC133+ or 5E12+ cells in the culture medium;
- (c) culturing the progeny of the isolated AC133+ or 5E12+ cell under conditions in which the isolated AC133+ cell or 5E12+ cell differentiates to neurons, astrocytes, and oligodendrocytes;
- and
- (d) detecting the presence of neurons, astrocytes, and oligodendrocytes, wherein the presence of neurons, astrocytes, and oligodendrocytes characterizes the isolated AC133+ cell or 5E12+ cell as an NS-IC.
A method for identifying the presence of a growth factor that affects the growth of a neurosphere initiating stem cell (NS-IC), the method comprising:
- (a) combining a composition suspected of containing at least one growth factor that affects the growth of NS-IC with a composition comprising NS-IC, wherein the NS-IC are characterized as AC133+ or 5E12+, nestin+, and capable of differentiation to neurons, astrocytes, and oligodendrocytes lineages;
- and
- (b) determining the growth of the NS-IC as a function of the presence of at least one of the growth factors in the composition, wherein an altered NS-IC growth, as compared with growth of NS-IC that have not been contacted with the composition suspected of containing at least one growth factor that affects the growth of NS-IC, indicates the presence in the composition of a growth factor that affects the growth of NS-IC.
Dependencies are inferred from the claim text.