Reprogramming cells
AssigneeThe Scripps Research Institute
InventorsSaiyong Zhu, Sheng Ding
Life of this patent
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Abstract
The present invention provides for methods, compositions, and kits for producing an induced pluripotent stem cell from a non-pluripotent mammalian cell using a 3′-phosphoinositide-dependent kinase-1 (PDK1) activator or a compound that promotes glycolytic metabolism as well as other small molecules.
Claims30
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A composition comprising an induced pluripotent stem cell (iPSC), wherein the iPSC is obtained by a method comprising:
- (a) introducing into a non-pluripotent cell a polynucleotide encoding an Oct4 polypeptide;
- (b) contacting the non-pluripotent cell with
- (i) a small molecule selected from the group consisting of F2,6P (fructose 2,6-bisphosphate), F6P (fructose 6-phosphate), DNP (2,4-dinitrophenol), NOG (N-oxalylglycine), QC (quercetin), 2-HA (2-hydroxyglutaric acid), NA (nicotinic acid), and PDK1 activator (3′-phosphoinositide-dependent kinase-1), and
- (ii) one or both of a TGFβ receptor/ALK5 inhibitor and a MEK inhibitor;
- and
- (c) culturing the contacted non-pluripotent cell of (b), thereby reprogramming the non-pluripotent mammalian cell to produce an induced pluripotent stem cell;
- wherein contacting the non-pluripotent cell with the small molecule enhances reprogramming when compared to without the small molecule.
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A therapeutic composition comprising an induced pluripotent stem cell (iPSC), wherein the iPSC is obtained by a method comprising:
- (a) introducing into a non-pluripotent cell a polynucleotide encoding an Oct4 polypeptide;
- (b) contacting the non-pluripotent cell with
- (i) a small molecule selected from the group consisting of F2,6P (fructose 2,6-bisphosphate), F6P (fructose 6-phosphate), DNP (2,4-dinitrophenol), NOG (N-oxalylglycine), QC (quercetin), 2-HA (2-hydroxyglutaric acid), NA (nicotinic acid), and PDK1 activator (3′-phosphoinositide-dependent kinase-1), and
- (ii) one or both of a TGFβ receptor/ALK5 inhibitor and a MEK inhibitor;
- and
- (c) culturing the contacted non-pluripotent cell of (b), thereby reprogramming the non-pluripotent mammalian cell to produce an induced pluripotent stem cell;
- wherein contacting the non-pluripotent cell with the small molecule enhances reprogramming when compared to without the small molecule.
A composition for use in manufacturing a pluripotent stem cell for application in regenerative medicine, wherein the composition comprises a small molecule, wherein the small molecule enhances reprogramming of a mammalian cell, and wherein the small molecule is selected from the group consisting of F2,6P (fructose 2,6-bisphosphate), F6P (fructose 6-phosphate), DNP (2,4-dinitrophenol), NOG (N-oxalylglycine), QC (quercetin), 2-HA (2-hydroxyglutaric acid), NA (nicotinic acid), or a PDK1 (3′-phosphoinositide-dependent kinase-1) activator.
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A kit comprising a small molecule that enhances reprogramming when used for a method of obtaining an induced pluripotent stem cell (iPSC) comprising:
- (a) introducing into a non-pluripotent cell a polynucleotide encoding an Oct4 polypeptide;
- (b) contacting the non-pluripotent cell with
- (i) the small molecule, wherein the small molecule enhancing reprogramming is selected from the group consisting of F2,6P (fructose 2,6-bisphosphate), F6P (fructose 6-phosphate), DNP (2,4-dinitrophenol), NOG (N-oxalylglycine), QC (quercetin), 2-HA (2-hydroxyglutaric acid), NA (nicotinic acid), and PDK1 activator (3′-phosphoinositide-dependent kinase-1), and
- (ii) a MEK inhibitor;
- and
- (c) culturing the contacted non-pluripotent cell of (b), thereby reprogramming the non-pluripotent mammalian cell to produce an induced pluripotent stem cell;
- wherein contacting the non-pluripotent cell with the small molecule enhances reprogramming when compared to without the small molecule.
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A method of making a therapeutic composition comprising an induced pluripotent stem cell (iPSC) for tissue regeneration in a subject in need thereof, wherein said iPSC is obtained by a method comprising:
- (a) introducing into a non-pluripotent cell a polynucleotide encoding an Oct4 polypeptide;
- (b) contacting the non-pluripotent cell with
- (i) a small molecule selected from the group consisting of F2,6P (fructose 2,6-bisphosphate), F6P (fructose 6-phosphate), DNP (2,4-dinitrophenol), NOG (N-oxalylglycine), QC (quercetin), 2-HA (2-hydroxyglutaric acid), NA (nicotinic acid), and PDK1 activator (3′-phosphoinositide-dependent kinase-1), and
- (ii) one or both of a TGFβ receptor/ALK5 inhibitor and a MEK inhibitor;
- and
- (c) culturing the contacted non-pluripotent cell of (b), thereby reprogramming the non-pluripotent mammalian cell to produce an induced pluripotent stem cell;
- wherein contacting the non-pluripotent cell with the small molecule enhances reprogramming when compared to without the small molecule.
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Dependencies are inferred from the claim text.