Method For Reducing Differentiation Resistance Of Pluripotent Stem Cells

Abstract

In related-art methods of differentiating pluripotent stem cells into a desired cell type, there has not been established a differentiation induction method using human ES/iPS cells and being stable and highly efficient. A method of inducing differentiation into a desired cell type within a short period of time and with high efficiency by attenuating differentiation resistance of a pluripotent stem cell to generate a pluripotent stem cell that actively proceeds to a differentiated cell type has been found, and thus the present invention has been completed.

1 .- 16 . (canceled) 
     
     
         17 . A differentiation induction kit for differentiating a pluripotent stem cell into a desired cell type, comprising at least any one of the following items (1) to (3):
 (1) a pluripotent stem cell having a histone in which H3K27me3 modification has been substantially removed or reduced;   (2) a pluripotent stem cell in which JMJD3 is forcibly expressed; and   (3) a pluripotent stem cell having a gene construct carrying JMJD3 inserted into a genome thereof,
 wherein the JMJD3 has demethylase activity that induces differentiation of the pluripotent stem cell by removing a methyl group of H3K27me3. 
   
     
     
         18 . The differentiation induction kit according to  claim 17 , wherein the JMJD3 is a demethylase containing only an enzymatically active region of JMJD3. 
     
     
         19 . The differentiation induction kit according to  claim 17 , wherein the JMJD3 has an amino acid sequence set forth in any one of SEQ ID NOS: 1 to 3. 
     
     
         20 . The differentiation induction kit according to  claim 17 , further comprising a transcription factor required for induction of differentiation into the desired cell type. 
     
     
         21 . The differentiation induction kit according to  claim 17 , wherein the desired cell type is a skeletal muscle cell, comprising at least any one of the following items (1) to (3):
 (1) a pluripotent stem cell having a histone in which H3K27me3 modification has been substantially removed or reduced, and a transcription factor MYOD1;   (2) a pluripotent stem cell in which JMJD3 is forcibly expressed, and a transcription factor MYOD1; and   (3) a pluripotent stem cell having a gene construct carrying JMJD3 inserted into a genome thereof, and a transcription factor MYOD1.   
     
     
         22 . The differentiation induction kit according to  claim 17 , wherein the desired cell type is a nerve cell, comprising at least any one of the following items (1) to (3):
 (1) a pluripotent stem cell having a histone in which H3K27me3 modification has been substantially removed or reduced, and a transcription factor NEUROG1, NEUROG2, NEUROG3, NEUROD1, and/or NEUROD2;   (2) a pluripotent stem cell in which JMJD3 is forcibly expressed, and a transcription factor NEUROG1, NEUROG2, NEUROG3, NEUROD1, and/or NEUROD2; and   (3) a pluripotent stem cell having a gene construct carrying JMJD3 inserted into a genome thereof, and a transcription factor NEUROG1, NEUROG2, NEUROG3, NEUROD1, and/or NEUROD2.   
     
     
         23 . The differentiation induction kit according to  claim 17 , wherein the desired cell type is a liver cell, comprising at least any one of the following items (1) to (3):
 (1) a pluripotent stem cell having a histone in which H3K27me3 modification has been substantially removed or reduced, and a transcription factor HNF1A;   (2) a pluripotent stem cell in which JMJD3 is forcibly expressed, and a transcription factor HNF1A; and   (3) a pluripotent stem cell having a gene construct carrying JMJD3 inserted into a genome thereof, and a transcription factor HNF1A.