Methods, kits, and compositions for stem cell self-renewal
The present invention relates to methods and kits for expanding a stem cell population. More particularly, the invention relates, inter alia, to methods, kits, and compositions for expanding a stem cell population, particularly a hematopoietic stem cell population.
1. An ex vivo method for expanding the number of hematopoietic stem cells (HSC) in a population of mononuclear cells (MNCs) comprising:
culturing the population of MNCs comprising at least one HSC in an HSC expansion media comprising a reversible glycogen synthase kinase 3 beta (GSK-3β) inhibitor for a period of time of at least 10 days, wherein the reversible GSK-3β inhibitor is effective to activate β-catenin, wherein the expanded HSCs are functional with long germ, multi-lineage, repopulation potential.
2. The method according to claim 1 , wherein the reversible GSK-3β inhibitor is selected from the group consisting of a small molecule, a biologic, an antisense RNA, a small interfering RNA (siRNA), and combinations thereof.
3. The method according to claim 2 , wherein the reversible GSK-3β inhibitor is a small molecule.
4. The method according to claim 2 , 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 1), Ro318220 (bisindolylmaleimide IX), SB216763, SB415286, 15, CGP60474, Compound 8b, TWS119, Compound 1A, 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.
5. The method according to claim 4 , wherein the reversible GSK-3β inhibitor is CHIR99021.
6. The method according to claim 1 , which provides HSCs that, upon transplant into a recipient, exhibit 60% to 100% donor repopulation at least 4 weeks after transplant into the recipient.
7. The method 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.
8. The method according to claim 1 , which provides HSCs that, upon transplant into a recipient, exhibit 5% to 100% donor repopulation at least 4 weeks after transplant into the recipient.
9. The method according to claim 1 , which provides HSCs that, upon transplant into a recipient, exhibit 25% to 100% donor repopulation at least 4 weeks after transplant into the recipient.
10. The method according to claim 1 , which provides HSCs that, upon transplant into a recipient, exhibit 45% to 100% donor repopulation at least 4 weeks after transplant into the recipient.