Method for rejuvenating cells

Abstract

The invention relates to a method for reprogramming cells from aged donors or senescent cells to pluripotent cells that have lost marks of senescence. In particular, the invention relates to an ex vivo method for preparing induced pluripotent stein cells (iPSCs) from a target cell population comprising cells from aged donors or senescent cells, said method comprising the steps of culturing said target cell population under appropriate conditions for reprogramming said cells into iPSCs, wherein said appropriate conditions comprises increasing expression in said target cells, of at least the following reprogramming factors: Oct4, Klf4, Sox2, Myc, Lin28 and, optionally Nanog.

1 - 15 . (canceled) 
     
     
         16 . Rejuvenated induced pluripotent stem cells (iPSCs) obtained by a method comprising the steps of:
 a) providing a cell composition obtained from an adult human subject that is at least 50 years old and,   b) culturing said cell composition under appropriate conditions for reprogramming said cell composition into rejuvenated iPSCs, wherein said appropriate conditions comprise increasing expression in said cell composition of at least the following combination of reprogramming factors:
 i. a reprogramming factor encoded by one gene of the Oct gene family, 
 ii. a reprogramming factor encoded by one gene of the Klf gene family, 
 iii. a reprogramming factor encoded by one gene of the Sox gene family, 
 iv. a reprogramming factor encoded by one gene of the Myc gene family, 
 v. Lin28, 
 vi. and Nanog, 
 thereby obtaining rejuvenated iPSC cells in which the expression of p21 CIP1  and p16 INK4A  is downregulated to a level found in human embryonic stem cells (hESC). 
   
     
     
         17 . The rejuvenated iPSCs of  claim 16 , wherein said combination of reprogramming factors comprises Oct4, Klf4, Sox2, c-Myc, Lin28 and Nanog. 
     
     
         18 . The rejuvenated iPSCs of  claim 16 , wherein said cell composition comprises or consists of senescent cells. 
     
     
         19 . The rejuvenated iPSCs of  claim 18 , wherein said senescent cells are obtained from a subject suffering from age-related disorders leading to a high proportion of senescent cells. 
     
     
         20 . The rejuvenated iPSCs of  claim 16 , wherein said method does not further comprise a step of silencing senescence effectors in said cell composition. 
     
     
         21 . The rejuvenated iPSCs of  claim 20 , wherein said senescence effectors include p21 CIP  and/or p16 INK4a  and/or p53. 
     
     
         22 . The rejuvenated iPSCs of  claim 16 , wherein said conditions for increasing expression of the reprogramming factors comprise
 a) introducing one or more expression vectors comprising coding sequences of said combination of reprogramming factors into said cell composition; or,   b) directly delivering an effective amount of each precursor RNA of the reprogramming factors into said cell composition.   
     
     
         23 . The rejuvenated iPSCs of  claim 22 , wherein said step of introducing is performed by transfecting said cell composition with said one or more expression vectors. 
     
     
         24 . Differentiated cells obtained by differentiation of the rejuvenated iPSCs of  claim 16  into a desired cell lineage.