Combined chemical and genetic 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 - 16 . (canceled) 
     
     
         17 . A therapeutic composition comprising:
 (a) an induced pluripotent stem (iPS) cell population, wherein the population comprises iPS cells reprogrammed from a mammalian non-pluripotent cell; and   (b) a medium comprising a MAPK/ERK kinase (MEK) inhibitor.   
     
     
         18 . The therapeutic composition of  claim 17 , wherein the iPS cell population has increased homogeneity in comparison to a iPS cell population without contacting the medium comprising a MEK inhibitor. 
     
     
         19 . The therapeutic composition of  claim 17 , wherein the medium promotes growth of iPS cells. 
     
     
         20 . The therapeutic composition of  claim 17 , wherein the medium inhibits growth of non-pluripotent cells. 
     
     
         21 . The therapeutic composition of  claim 17 , wherein MEK inhibitor is PD0325901. 
     
     
         22 . The therapeutic composition of  claim 17 , wherein the cell population is obtained by introducing one or more expression cassettes comprising a polynucleotide encoding an Oct3/4 polypeptide and a polynucleotide encoding a Klf polypeptide into non-pluripotent mammalian cells, thereby initiating reprogramming of the non-pluripotent cells. 
     
     
         23 . The therapeutic composition of  claim 22 , wherein the one or more expression cassettes further comprise a polynucleotide encoding a Sox polypeptide or a polynucleotide encoding a c-Myc polypeptide. 
     
     
         24 . The therapeutic composition of  claim 17 , wherein the non-pluripotent mammalian cells are somatic cells. 
     
     
         25 . The therapeutic composition of  claim 17 , wherein the non-pluripotent mammalian cells are progenitor cells. 
     
     
         26 . The therapeutic composition of  claim 17 , wherein the medium further comprises a GSK3 inhibitor, an Erk inhibitor, or a TGFβ receptor/ALK5 inhibitor. 
     
     
         27 . The therapeutic composition of  claim 17 , further comprising a pharmaceutically acceptable carrier. 
     
     
         28 . A composition comprising differentiated cells of a desired cell type derived from induced pluripotent stem (iPS) cellls, wherein the iPS cells are comprised in a medium comprising a MEK inhibitor. 
     
     
         29 . A therapeutic composition comprising differentiated cells according to  claim 28 . 
     
     
         30 . A method of using the therapeutic composition of  claim 17  or  29  by administering the composition to a subject suitable for such treatment. 
     
     
         31 . The method of  claim 30 , wherein the therapeutic composition comprises cells that are allogenic or autogenic. 
     
     
         32 . A kit comprising (i) an induced pluripotent stem cell reprogrammed from a mammalian non-pluripotent cell, and (ii) a medium comprising a MEK inhibitor. 
     
     
         33 . A kit comprising differentiated cells of a desired cell type derived from induced pluripotent stem (iPS) cellls, wherein the iPS cells are comprised in a composition comprising a medium comprising a MEK inhibitor.