Induced pluripotent stem cell selection method and method for inducing differentiation to blood cells
First Claim
1. A method for producing hematopoietic stem cells and/or hematopoietic progenitor cells from pluripotent stem (PS) cells, said method comprising:
- (a) measuring the expression level(s) of one or more genes selected from the group consisting of TRIM58, CTSF, FAM19A5, and TCERG1L genes in candidate PS cells;
(b) selecting a PS cell(s) in which the expression level(s) of the gene(s) measured in Step (a) is/are equivalent to or higher than the expression level(s) in iPS cells or embryonic stem (ES) cells which are known to have high capacity to differentiate into blood cells, and/or selecting a PS cell(s) in which the expression level(s) of the gene(s) measured in Step (a) is/are higher than the expression level(s) in iPS cells or ES cells which are known to have low capacity to differentiate into blood cells; and
(c) culturing the selected PS cells in the presence of insulin-like growth factor 2 (IGF2).
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Abstract
A method for producing hematopoietic stem cells and/or hematopoietic progenitor cells from pluripotent stem cells is described. The method includes a step of culturing pluripotent stem cells in the presence of IGF2. A method is described for selecting an induced pluripotent stem cell(s) having high capacity to differentiate into hematopoietic stem cells and/or hematopoietic progenitor cells, or into blood cells, based on the expression level(s) of one or more genes such as TRIM58, CTSF, FAM19A5, and TCERG1L genes, or on the DNA methylation state(s) of the TRIM58, CSMD1, and/or FAM19A5 gene(s).
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Citations
17 Claims
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1. A method for producing hematopoietic stem cells and/or hematopoietic progenitor cells from pluripotent stem (PS) cells, said method comprising:
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(a) measuring the expression level(s) of one or more genes selected from the group consisting of TRIM58, CTSF, FAM19A5, and TCERG1L genes in candidate PS cells; (b) selecting a PS cell(s) in which the expression level(s) of the gene(s) measured in Step (a) is/are equivalent to or higher than the expression level(s) in iPS cells or embryonic stem (ES) cells which are known to have high capacity to differentiate into blood cells, and/or selecting a PS cell(s) in which the expression level(s) of the gene(s) measured in Step (a) is/are higher than the expression level(s) in iPS cells or ES cells which are known to have low capacity to differentiate into blood cells; and (c) culturing the selected PS cells in the presence of insulin-like growth factor 2 (IGF2). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for producing hematopoietic stem cells and/or hematopoietic progenitor cells from pluripotent stem (PS) cells, said method comprising
(a) measuring the DNA methylation state(s) of TRIM58, CSMD1, and/or FAM19A5 gene(s) in candidate PS cells; -
(b) selecting a PS cell(s) in which the DNA methylation level(s) measured in Step (a) is/are equivalent to or lower than the DNA methylation level(s) in iPS cells or ES cells which are known to have high capacity to differentiate into blood cells, and/or selecting a PS cell(s) in which the DNA methylation level(s) measured in Step (a) is/are lower than the DNA methylation level(s) in iPS cells or ES cells which are known to have low capacity to differentiate into blood cells; and (c) culturing the selected PS cells in the presence of insulin-like growth factor 2 (IGF2). - View Dependent Claims (11, 12, 13, 14, 15, 16, 17)
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Specification