Chemical approaches for generation of induced pluripotent stem cells

Abstract

The present invention provides for identification and use of small molecules to induce pluripotency in mammalian cells as well as other methods of inducing pluripotency.

1 - 20 . (canceled) 
     
     
         21 . A therapeutic composition comprising (i) an induced pluripotent stem cell reprogrammed from a mammalian non-pluripotent cell, and (ii) a small molecule composition comprising a GSK-3 inhibitor. 
     
     
         22 . The therapeutic composition of  claim 21 , wherein the small molecule composition further comprises a MEK inhibitor, an Erk inhibitor, or a TGFβ receptor/ALK5 inhibitor. 
     
     
         23 . The therapeutic composition of  claim 22 , wherein the TGFβ receptor/ALK5 inhibitor is selected from the group consisting of A-83-01 and SB431542. 
     
     
         24 . The therapeutic composition of  claim 22 , wherein the MEK inhibitor is PD0325901. 
     
     
         25 . The therapeutic composition of  claim 21 , wherein the GSK-3 inhibitor is selected from the group consisting of CHIR99021, CHIR98014, and 6-bromoindirubin-3′-oxime (BIO). 
     
     
         26 . The therapeutic composition of  claim 21 , wherein the induced pluripotent stem cell is reprogrammed by introducing into the mammalian non-pluripotent cell exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf encoding polynucleotide to initiate reprogramming. 
     
     
         27 . The therapeutic composition of  claim 21 , further comprising a pharmaceutically acceptable carrier. 
     
     
         28 . A composition comprising differentiated cells of a desired cell type derived from induced pluripotent stem (iPS) cells, wherein the iPS cells are comprised in a composition comprising a small molecule composition comprising a GSK-3 inhibitor. 
     
     
         29 . The composition of  claim 28 , wherein the GSK-3 inhibitor is selected from the group consisting of CHIR99021, CHIR98014, and 6-bromoindirubin-3′-oxime (BIO). 
     
     
         30 . The composition of  claim 28 , wherein the iPS cells are reprogrammed by introducing into mammalian non-pluripotent cells exogenous polynucleotides comprising an Oct4 encoding polynucleotide and a Klf encoding polynucleotide to initiate reprogramming. 
     
     
         31 . A therapeutic composition comprising differentiated cells according to  claim 28 . 
     
     
         32 . A method of using the therapeutic composition of  claim 21  by administering the composition to a subject suitable for treatment. 
     
     
         33 . The method of  claim 32 , wherein the therapeutic composition comprises cells that are allogenic or autogenic. 
     
     
         34 . A method of using the therapeutic composition of  claim 31  by administering the composition to a subject suitable for treatment. 
     
     
         35 . The method of  claim 34 , wherein the therapeutic composition comprises cells that are allogenic or autogenic. 
     
     
         36 . A kit comprising (i) an induced pluripotent stem cell reprogrammed from a mammalian non-pluripotent cell, and (ii) a small molecule composition comprising a GSK-3 inhibitor. 
     
     
         37 . A kit comprising differentiated cells of a desired cell type derived from induced pluripotent stem (iPS) cells, wherein the iPS cells are comprised in a composition comprising a small molecule composition comprising a GSK-3 inhibitor.