Compositions of enriched central nervous system stem cell and progenitor cell populations
View Patent ↗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.
1. A composition comprising:
(a) a population of neural cells enriched for neurosphere initiating stem cells (NS-IC) produced by:
(1) combining a population comprising neural cells or neural-derived cells containing a fraction of NS-ICs with monoclonal antibody AC133 or monoclonal antibody 5E12 (ATCC Accession No. PTA-994); and
(2) selecting and isolating those cells that bind to monoclonal antibody AC133 or monoclonal antibody 5E12, thereby producing a population of cells enriched for NS-IC as compared with the population of neural cells or neural-derived cells; and
(b) at least one monoclonal antibody selected from the group consisting of monoclonal antibody AC133 and monoclonal antibody 5E12, wherein the at least one monoclonal antibody is conjugated with a label to allow for ease of separation.
2. An in vitro cell culture composition comprising:
(a) a population of neural cells enriched in NS-IC, wherein the NS-IC bind to monoclonal antibody AC133 or to monoclonal antibody 5E12 (ATCC Accession No. PTA-994) and are CD45 − cells;
(b) a medium capable of supporting the growth of the cells; and
(c) at least one monoclonal antibody selected from the group consisting of monoclonal antibody AC133 and monoclonal antibody 5E12 , wherein the at least one monoclonal antibody is conjugated with a label to allow for ease of separation.
3. An in vitro cell culture composition comprising:
(a) a population of neural cells enriched in NS-IC, wherein the NS-IC bind to monoclonal antibody AC133 or to monoclonal antibody 5E12 (ATCC Accession No. PTA-994) and are CD45 − CD34 − cells;
(b) a medium capable of supporting the growth of the cells; and
(c) at least one monoclonal antibody selected from the group consisting of monoclonal antibody AC133 and monoclonal antibody 5E12, wherein the at least one monoclonal antibody is conjugated with a label to allow for ease of separation.
4. An in vitro cell culture composition comprising:
(a) a population of neural cells enriched in NS-IC, wherein the NS-IC bind to monoclonal antibody AC133 or to monoclonal antibody 5E12 (ATCC Accession No. PTA-994) and are CD24 − cells;
(b) a medium capable of supporting the growth of the cells; and
(c) at least one monoclonal antibody selected from the group consisting of monoclonal antibody AC133 and monoclonal antibody 5E12, wherein the at least one monoclonal antibody is conjugated with a label to allow for ease of separation.
5. The composition of any one of claims 1 and 2 – 4 , further comprising a solid support to which the cells are attached.
6. The composition of any one of claims 1 and 2 – 4 , wherein the population of cells comprises at least 70% cells that bind to monoclonal antibody AC133 or monoclonal antibody 5E12.
7. The composition of any one of claims 1 and 2 – 4 , wherein the population of cells comprises at least 90% cells that bind to monoclonal antibody AC133 or monoclonal antibody 5E12.
8. The composition of any one of claims 1 and 2 – 4 , wherein the population of cells that bind to monoclonal antibody AC133 or monoclonal antibody 5E12 is a substantially pure population.
9. The composition of any one of claims 1 and 2 – 4 , wherein the medium comprises a serum-free medium containing one or more growth factors effective for inducing multipotent neural stem cell proliferation.
10. The composition of any one of claims 1 and 2 – 4 , wherein the medium comprises a growth factor selected from the group consisting of leukocyte inhibitory factor (LIF), epidermal growth factor (EGF), basic fibroblast growth factor (FGF-2), and combinations thereof.
11. The composition of any one of claims 1 and 2 – 4 , wherein the medium comprises neural survival factor, NSF.
12. The composition of any one of claims 1 and 2 – 4 , wherein the neural cells are human.
13. The composition of any one of claims 1 and 2 – 4 , wherein the label is selected from the group consisting of magnetic beads, magnetic reagents, superparamagnetic microparticles, biotin, fluorochromes, and haptens.