Media for stem cell proliferation and induction
The present application discloses a cell culture media for growth, maintenance and induction of reversion to a less mature state of a cell comprising a MUC1* activating ligand.
1. A method for stimulating growth of stem or progenitor cells or for inducing cells to revert to a less mature state, the method comprising contacting cells with cell culture media that comprises a recombinant NME7 protein that comprises a nucleotide diphosphate kinase (NDPK) A domain and an NDPK B domain and does not comprise a targeting sequence.
2. The method according to claim 1 , wherein the stem or progenitor cells are grown or the cells are induced to revert to the less mature state on a surface coated with anti-MUC1* antibody.
3. The method according to claim 1 , wherein the recombinant NME7 is the only growth factor in the cell culture media.
4. The method according to claim 1 , wherein the cell culture media does not comprise basic fibroblast growth factor (bFGF) and transforming growth factor beta (TGF-beta).
5. The method according to claim 1 , wherein the cells are mature, somatic or progenitor cells.
6. The method according to claim 1 , wherein the cells are human cells.
7. The method according to claim 1 , wherein the cell culture media is free of serum.
8. The method according to claim 1 , wherein the cell culture media further comprises insulin, selenium, transferrin, L ascorbic acid, or one or more non-essential amino acids.
9. The method according to claim 1 , wherein the recombinant NME7 is characterized by a molecular weight of about 25 to 33 kilodaltons (kDa).
10. The method according to claim 1 , wherein the recombinant NME7 protein is characterized by a molecular weight of about 30 kilodaltons (kDa).
11. The method according to claim 1 , wherein the recombinant NME7 has a molecular weight of about 31 kilodaltons (kDa).
12. The method according to claim 1 , wherein the recombinant NME7 is characterized by a molecular weight of about 33 kilodaltons (kDa).
13. The method according to claim 1 , wherein the recombinant NME7 comprises the amino acid sentience of SEQ ID NO: 39 41 59 or 61.
14. The method according to claim 1 , wherein the recombinant NME7 comprises an alternative splice isoform having an amino acid sequence of SEQ ID NO: 39, 41, 59 or 61.
15. The method according to claim 1 , wherein the cell culture media further comprises an inhibitor of a rho associated kinase.
16. The method according to claim 1 , wherein the cell culture media further comprises an activator of one or more signaling proteins in the PBK or RAC pathway.
17. The method according to claim 1 , wherein the cell culture media further comprises an activator of one or more signaling proteins in the PBK or RAC pathway in the absence of a rho kinase inhibitor.
18. The method according to claim 1 , wherein the cell culture media further comprises a nucleic acid that suppresses expression of NME1 or NME2.
19. The method according to claim 1 , wherein the recombinant NME7 has superior high yield expression of the soluble, properly folded protein in E. coli compared with native NME7.