Methods and compositions for stem cell self-renewal
The present invention relates to methods for expanding a stem cell population. More particularly, the invention relates, inter alia, to methods and compositions for expanding a stem cell population, particularly a hematopoietic stem cell population.
1. A media for carrying out ex vivo expansion of a hematopoietic stem cell (HSC) population comprising a fluid media suitable for maintaining viable stem cells and PTEN and GSK-3β inhibitors present in the media at concentrations sufficient to enable expansion of the HSC population while maintaining a multilineage differentiation potential in the HSCs.
2. The media according to claim 1 , wherein the expansion of the number of stem cells is by a factor selected from the group consisting of at least 40-fold, at least 80-fold, at least 150-fold, and at least 250-fold.
3. The media according to claim 1 , wherein the HSC is obtained from a mammalian tissue selected from the group consisting of cord blood, peripheral blood, and bone marrow.
4. The media according to claim 1 , wherein the PTEN inhibitor is reversible.
5. The media according to claim 4 , wherein the reversible PTEN inhibitor is selected from the group consisting of shikonin, a bisperoxovanadium compound, SF-1751 (Semafore Pharmaceuticals), pharmaceutical salts thereof, and combinations thereof.
6. The media according to claim 5 , wherein the bisperoxovanadium compound is selected from the group consisting of bpV(phen)2, bpV(pic), pharmaceutical salts thereof, and combinations thereof.
7. The media according to claim 1 wherein the GSK-3β inhibitor is reversible.
8. The media according to claim 7 , wherein the reversible GSK-3β inhibitor is selected from the group consisting of Hymenialdisine, Flavopiridol, Kenpaullone, Alsterpaullone, Azakenpaullone, Indirubin-30-oxime, 6-Bromoindirubin-30-oxime (BIO), 6-Bromoindirubin-30-acetoxime, Aloisine A, Aloisine B, TDZD8, Compound 12, CHIR98014, CHIR99021 (CT99021), CT20026, Compound 1, SU9516, ARA014418, Staurosporine, Compound 5a, Compound 29, Compound 46, GF109203x (bisindolylmaleimide I), Ro318220 (bisindolylmaleimide IX), SB216763, SB415286, 15, CGP60474, Compound 8b, TWS119, Compound IA, Compound 17, Lithium, Beryllium, Zinc, small molecule GSK-3β inhibitors (Vertex Pharmaceuticals), NP-12 (Neuropharma), GSK-3β inhibitors (Amphora), GSK-3β inhibitors (CrystalGenomics), SAR-502250 (Sanofi-Aventis), 3544 (Hoffmann-La Roche), GSK-3β inhibitors (Lundbeck), TDZD-8 (Cancer Center, University of Rochester), pharmaceutically acceptable salts thereof, and combinations thereof.
9. The media according to claim 8 , wherein the reversible GSK-3β inhibitor is CHIR99021.
10. The media according to claim 1 , wherein the PTEN inhibitor is bpV(pic), and the GSK-3β inhibitor is CHIR99021.
11. The media according to claim 2 , wherein the expansion of the number of stem cells is by a factor of at least 40-fold.
12. The media according to claim 2 , wherein the expansion of the number of stem cells is by a factor of at least 80-fold.
13. The media according to claim 2 , wherein the expansion of the number of stem cells is by a factor of at least 150-fold.
14. The media according to claim 2 , wherein the expansion of the number of stem cells is by a factor of at least 250-fold.