Method of efficiently establishing induced pluripotent stem cells

Abstract

The invention provides an in vitro method of improving the efficiency of establishment of induced pluripotent stem (iPS) cells. The method comprises contacting an isolated somatic cell being reprogrammed into an iPS cell with an inhibitor of p53 function. The invention also provides an in vitro method of producing iPS cells. The method comprises bringing (a) nuclear reprogramming substances or nucleic acids encoding the nuclear reprogramming substances and (b) an inhibitor of p53 function into contact with a somatic cell.

1 . An in vitro method of improving the efficiency of establishment of an induced pluripotent stem (iPS) cell, comprising contacting an isolated somatic cell being reprogrammed into an iPS cell with an inhibitor of p53 function, wherein the inhibitor of p53 function is selected from the following:
 (i) a dominant negative mutant of p53 or a nucleic acid that encodes the same,   (ii) PFT-α or an analogue thereof, PFT-β or an analogue thereof, or PFT-μ,   (iii) MDM2 or a nucleic acid that encodes the same, and   (iv) an siRNA, shRNA, antisense nucleic acid or ribozyme against p21, or an anti-p21 antagonist antibody or a nucleic acid that encodes the same.   
     
     
         2 . The method of  claim 1 , wherein the inhibitor of p53 function is a dominant negative mutant of p53 or a nucleic acid that encodes the same. 
     
     
         3 . The method of  claim 2 , wherein the dominant negative mutant of p53 is p53DD, p53P275S or p53D278N. 
     
     
         4 . The method of  claim 2 , wherein the dominant negative mutant of p53 is p53DD. 
     
     
         5 . An in vitro method of producing iPS cells, comprising bringing (a) nuclear reprogramming substances or nucleic acids encoding the nuclear reprogramming substances and (b) an inhibitor of p53 function into contact with a somatic cell, wherein the nuclear reprogramming substances are (i) Oct3/4 and Klf4, (ii) Oct3/4 and c-Myc, (iii) Oct3/4, Klf4 and Sox2, (iv) Oct3/4, Klf4 and c-Myc, or (v) Oct3/4, Klf4, Sox2 and c-Myc, and wherein the inhibitor of p53 function is selected from the following:
 (i) a dominant negative mutant of p53 or a nucleic acid that encodes the same,   (ii) PFT-α or an analogue thereof, PFT-β or an analogue thereof, or PFT-μ,   (iii) MDM2 or a nucleic acid that encodes the same, and   (iv) an siRNA, shRNA, antisense nucleic acid or ribozyme against p21, or an anti-p21 antagonist antibody or a nucleic acid that encodes the same.   
     
     
         6 . The method of  claim 5 , wherein the inhibitor of p53 function is a dominant negative mutant of p53 or a nucleic acid that encodes the same. 
     
     
         7 . The method of  claim 6 , wherein the dominant negative mutant of p53 is p53DD, p53P275S or p53D278N. 
     
     
         8 . The method of  claim 6 , wherein the dominant negative mutant of p53 is p53DD. 
     
     
         9 . The method of  claim 5 , wherein the nuclear reprogramming substances are Oct3/4, Klf4 and Sox2, or nucleic acids that encode the same. 
     
     
         10 . The method of  claim 5 , wherein the nuclear reprogramming substances are Oct3/4, Klf4, Sox2 and c-Myc, or nucleic acids that encode the same. 
     
     
         11 . The method of  claim 5 , wherein the somatic cell is a T cell.