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1.发明申请公开(公告)号：US20200370020A1
公开(公告)日：2020-11-26
申请号：US16988145
申请日：2020-08-07
IPC分类号： C12N5/071
摘要： An object of the present invention is to provide a new means capable of easily and efficiently preparing cells showing a function more similar to the function of intestinal epithelial cells of a living body. According to the present invention, pluripotent stem cells are induced to differentiate into intestinal stem cell-like cells by a method for inducing differentiation of pluripotent stem cells into intestinal epithelial cells, the method including a step (1) of differentiating pluripotent stem, cells into intestinal stem cell-like cells and a step (2) of differentiating the intestinal stem cell-like cells obtained in the step (1) into intestinal epithelial cell-like cells by using an MEK1 inhibitor, a DNA methylation inhibitor, a TGFβ receptor inhibitor, EGF, and a cAMP activator in combination.
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2.发明授权公开(公告)号：US12054746B2
公开(公告)日：2024-08-06
申请号：US16988145
申请日：2020-08-07
IPC分类号： C12N5/071
CPC分类号： C12N5/0679 , C12N2501/11 , C12N2501/115 , C12N2501/999 , C12N2506/45
摘要： An object of the present invention is to provide a new means capable of easily and efficiently preparing cells showing a function more similar to the function of intestinal epithelial cells of a living body. According to the present invention, pluripotent stem cells are induced to differentiate into intestinal stem cell-like cells by a method for inducing differentiation of pluripotent stem cells into intestinal epithelial cells, the method including a step (1) of differentiating pluripotent stem, cells into intestinal stem cell-like cells and a step (2) of differentiating the intestinal stem cell-like cells obtained in the step (1) into intestinal epithelial cell-like cells by using an MEK1 inhibitor, a DNA methylation inhibitor, a TGFβ receptor inhibitor, EGF, and a cAMP activator in combination.
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公开(公告)号：US11725189B2
公开(公告)日：2023-08-15
申请号：US16544993
申请日：2019-08-20
发明人： Takahiro Iwao , Tamihide Matsunaga , Satoshi Kondo , Shota Mizuno
CPC分类号： C12N5/0679 , G01N33/5014 , G01N33/5044 , C12N2501/115 , C12N2501/119 , C12N2501/415 , C12N2501/999 , C12N2506/45
摘要： It is an object to provide a culture method which is capable of maintaining and/or culturing iPS cell-derived intestinal stem cells, while maintaining the properties of intestinal stem cells. The induced pluripotent stem cell-derived intestinal stem cell-like cells are cultured in the presence of a GSK-3β inhibitor, a histone deacetylation inhibitor, and a serum replacement, or in the presence of a GSK-3β inhibitor and a serum replacement. Preferably, the culture is carried out under conditions in which one or more compounds selected from the group consisting of an epidermal growth factor, a TGFβ receptor inhibitor and a fibroblast growth factor are further present.
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4.发明申请公开(公告)号：US20190079076A1
公开(公告)日：2019-03-14
申请号：US16083226
申请日：2017-03-03
发明人： Takahiro Iwao , Tomoki Kabeya , Tamihide Matsunaga
摘要： An object of the present invention is to provide a novel method which enables convenient preparation of cells exhibiting functions close to that of intestinal epithelial cells of living bodies, and use of the method. The differentiation of induced pluripotent stem cells into intestinal epithelial cells is induced by step of differentiating induced pluripotent stem cells into endoderm-like cells; step of differentiating the endoderm-like cells obtained in step into intestinal stem cell-like cells; and step of differentiating the intestinal stem cell-like cells obtained in step into intestinal epithelial cell-like cells, wherein step includes culture in the presence of a MEK1 inhibitor, a DNA methyltransferase inhibitor, a TGF-β receptor inhibitor, and EGF and under the condition that cAMP is supplied to the cells.
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公开(公告)号：US20200157507A1
公开(公告)日：2020-05-21
申请号：US16611359
申请日：2018-05-02
发明人： Tamihide Matsunaga , Takahiro Iwao , Daichi Onozato , Isamu Ogawa
摘要： An object of the present invention is to prepare a functional intestinal organoid from pluripotent stem cells. An intestinal organoid is prepared from pluripotent stem cells, by the following steps (1) to (4): (1) differentiating pluripotent stem cells into endoderm-like cells; (2) differentiating the endoderm-like cells obtained in step (1) into intestinal stem cell-like cells; (3) culturing the intestinal stem cell-like cells obtained in step (2) to form spheroids; and (4) differentiating the spheroids formed in step (3) to form an intestinal organoid, the step including culture in the presence of a MEK1/2 inhibitor, a DNA methylation inhibitor, a TGF-β receptor inhibitor, and a γ-secretase inhibitor, in addition to an epidermal growth factor, a BMP inhibitor, and a Wnt signal activator.
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公开(公告)号：US11859212B2
公开(公告)日：2024-01-02
申请号：US16611359
申请日：2018-05-02
发明人： Tamihide Matsunaga , Takahiro Iwao , Daichi Onozato , Isamu Ogawa
CPC分类号： C12N5/0679 , A61L27/3834 , A61L27/3882 , A61L27/3895 , G01N33/5082 , C12N2501/11 , C12N2501/115 , C12N2501/119 , C12N2501/998 , C12N2501/999 , C12N2503/04 , C12N2506/45 , C12N2513/00 , C12N2533/70
摘要： An object of the present invention is to prepare a functional intestinal organoid from pluripotent stem cells. An intestinal organoid is prepared from pluripotent stem cells, by the following steps (1) to (4): (1) differentiating pluripotent stem cells into endoderm-like cells; (2) differentiating the endoderm-like cells obtained in step (1) into intestinal stem cell-like cells; (3) culturing the intestinal stem cell-like cells obtained in step (2) to form spheroids; and (4) differentiating the spheroids formed in step (3) to form an intestinal organoid, the step including culture in the presence of a MEK1/2 inhibitor, a DNA methylation inhibitor, a TGF-β receptor inhibitor, and a γ-secretase inhibitor, in addition to an epidermal growth factor, a BMP inhibitor, and a Wnt signal activator.
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7.发明授权公开(公告)号：US11499963B2
公开(公告)日：2022-11-15
申请号：US16083226
申请日：2017-03-03
发明人： Takahiro Iwao , Tomoki Kabeya , Tamihide Matsunaga
摘要： An object of the present invention is to provide a novel method which enables convenient preparation of cells exhibiting functions close to that of intestinal epithelial cells of living bodies, and use of the method. The differentiation of induced pluripotent stem cells into intestinal epithelial cells is induced by step of differentiating induced pluripotent stem cells into endoderm-like cells; step of differentiating the endoderm-like cells obtained in step into intestinal stem cell-like cells; and step of differentiating the intestinal stem cell-like cells obtained in step into intestinal epithelial cell-like cells, wherein step includes culture in the presence of a MEK1 inhibitor, a DNA methyltransferase inhibitor, a TGF-β receptor inhibitor, and EGF and under the condition that cAMP is supplied to the cells.
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