Method For Reducing Differentiation Resistance Of Pluripotent Stem Cells
AssigneeKeio University
InventorsMinoru Ko, Tomohiko Akiyama
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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.
Claims14
1-16. (canceled)
A method of differentiating a pluripotent stem cell into a desired cell type, comprising any one of the following steps (1) to (7):
- (1) a step of adding a demethylase gene and a transcription factor required for induction of differentiation into the desired cell type to a pluripotent stem cell;
- (2) a step of inserting a gene construct carrying a demethylase gene and a transcription factor gene required for induction of differentiation into the desired cell type into a genome of a pluripotent stem cell;
- (3) a step of inserting a gene construct carrying a demethylase gene into a genome of a pluripotent stem cell, followed by addition of a transcription factor required for induction of differentiation into the desired cell type to the cell;
- (4) a step of inserting a gene construct carrying a demethylase gene and a gene construct carrying a transcription factor required for induction of differentiation into the desired cell type into a genome of a pluripotent stem cell;
- (5) a step of adding a transcription factor required for induction of differentiation into the desired cell type to a pluripotent stem cell having a histone in which H3K27me3 modification has been substantially removed or reduced;
- (6) a step of adding a transcription factor required for induction of differentiation into the desired cell type to a pluripotent stem cell in which a demethylase is forcibly expressed;
- and
- (7) a step of adding a demethylase and a transcription factor required for differentiation into the desired cell type to a pluripotent stem cell.
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 (5):
- (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 a demethylase is forcibly expressed;
- (3) a pluripotent stem cell and a demethylase gene;
- (4) a gene construct carrying a demethylase gene and a pluripotent stem cell;
- and
- (5) a pluripotent stem cell having a gene construct carrying a demethylase gene inserted into a genome thereof.
A differentiation induction kit for differentiating a pluripotent stem cell into a skeletal muscle cell, comprising at least any one of the following items (1) to (5):
- (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 a demethylase is forcibly expressed, and a transcription factor MYOD1;
- (3) a pluripotent stem cell, a demethylase gene, and a transcription factor MYOD1;
- (4) a gene construct carrying a demethylase gene, a pluripotent stem cell, and a transcription factor MYOD1;
- and
- (5) a pluripotent stem cell having a gene construct carrying a demethylase gene inserted into a genome thereof, and a transcription factor MYOD1.
Dependencies are inferred from the claim text.