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公开(公告)号：US20220119764A1
公开(公告)日：2022-04-21
申请号：US17567708
申请日：2022-01-03
发明人： Atsushi KUWAHARA , Suguru YAMASAKI , Yasushi HIRAMINE , Yoshiki SASAI (deceased) , Masayo TAKAHASHI
IPC分类号： C12N5/079 , A61K35/44 , A61L27/00 , C12Q1/02 , C12N5/0797 , A61K35/30 , A61L27/38 , C12N5/0793 , G01N33/50
摘要： The present invention provides a method for producing retinal cells or a retinal tissue, comprising the following steps (1)-(3): (1) a first step of culturing human pluripotent stem cells in the absence of feeder cells and in a medium comprising a factor for maintaining undifferentiated state, (2) a second step of culturing the pluripotent stem cells obtained in the first step in suspension in the presence of a Sonic hedgehog signal transduction pathway activating substance to form a cell aggregate, and (3) a third step of culturing the aggregate obtained in the second step in suspension in the presence of a 1) a BMP signal transduction pathway activating substance to obtain an aggregate containing retinal cells or a retinal tissue.
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公开(公告)号：US20190127690A1
公开(公告)日：2019-05-02
申请号：US16095339
申请日：2017-04-21
IPC分类号： C12N5/074 , A61K35/30 , A61P27/06 , A61P9/10 , G01N33/15 , A61L27/38 , A61L27/54 , G01N33/50
摘要： The present invention provides a method for producing a retinal cell or a retinal tissue, including the following steps (1)-(3): (1) a first step of culturing pluripotent stem cells in the absence of feeder cells in a medium containing 1) a TGFβ family signal transduction pathway inhibiting substance and/or a Sonic hedgehog signal transduction pathway activating substance, and 2) a factor for maintaining undifferentiated state, (2) a second step of culturing the cells obtained in the first step in suspension in a medium containing a Wnt signal transduction pathway inhibiting substance to form a cell aggregate, and (3) a third step of culturing the aggregate obtained in the second step in suspension in the presence or absence of a Wnt signal transduction pathway inhibiting substance in a medium containing a BMP signal transduction pathway activating substance to obtain an aggregate containing a retinal cell or a retinal tissue.
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公开(公告)号：US20200208103A1
公开(公告)日：2020-07-02
申请号：US16632362
申请日：2018-07-20
发明人： Hideya SAKAGUCHI , Yoshiki SASAI (deceased) , Mototsugu EIRAKU , Daiki NUKAYA , Atsushi KUWAHARA
IPC分类号： C12N5/0793
摘要： The present invention provides a method for maintaining a continuous epithelial structure of a retinal tissue including culturing the retinal tissue in a medium comprising a methyl group donor or a substrate of the methyl group donor at a concentration at which cell differentiation of a neural retinal progenitor cell is suppressed, and a neurite extension inhibitor at a concentration at which neurite extension is suppressed.
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公开(公告)号：US20170313976A1
公开(公告)日：2017-11-02
申请号：US15521334
申请日：2015-10-23
IPC分类号： C12N5/0793 , A61K35/30 , C12N5/079 , G01N33/50 , A61L27/38
CPC分类号： C12N5/062 , A61K35/12 , A61K35/30 , A61L27/00 , A61L27/383 , A61L27/3895 , C12N5/0618 , C12N5/0619 , C12N2500/99 , C12N2501/115 , C12N2501/15 , C12N2501/155 , C12N2501/16 , C12N2501/40 , C12N2501/41 , C12N2501/415 , C12N2501/727 , C12N2506/45 , C12N2533/52 , C12Q1/02 , G01N33/5014
摘要： The present invention provides a method for producing neural cells or a neural tissue, including the following steps (1)-(3): (1) a first step of culturing pluripotent stem cells in the absence of feeder cells and in a medium containing 1) a TGFβ family signal transduction pathway inhibiting substance and/or a Sonic hedgehog signal transduction pathway activating substance, and 2) a factor for maintaining undifferentiated state, (2) a second step of culturing the cells obtained in the first step in suspension to form a cell aggregate, and (3) a third step of culturing the aggregate obtained in the second step in suspension in the presence or absence of a differentiation-inducing factor to obtain an aggregate containing neural cells or a neural tissue.
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公开(公告)号：US20220112457A1
公开(公告)日：2022-04-14
申请号：US17559458
申请日：2021-12-22
发明人： Atsushi KUWAHARA , Suguru YAMASAKI , Yasushi HIRAMINE , Yoshiki SASAI (deceased) , Masayo TAKAHASHI
IPC分类号： C12N5/0793 , A61K35/30 , A61L27/00 , A61L27/38 , C12N5/079 , C12N5/0797 , C12Q1/02 , G01N33/50 , A61K35/12
摘要： The present invention provides a method for producing neural cells or a neural tissue, including the following steps (1)-(3):
(1) a first step of culturing pluripotent stem cells in the absence of feeder cells and in a medium containing 1) a TGFβ family signal transduction pathway inhibiting substance and/or a Sonic hedgehog signal transduction pathway activating substance, and 2) a factor for maintaining undifferentiated state, (2) a second step of culturing the cells obtained in the first step in suspension to form a cell aggregate, and (3) a third step of culturing the aggregate obtained in the second step in suspension in the presence or absence of a differentiation-inducing factor to obtain an aggregate containing neural cells or a neural tissue.-
公开(公告)号：US20170313981A1
公开(公告)日：2017-11-02
申请号：US15521387
申请日：2015-10-23
IPC分类号： C12N5/079 , A61L27/38 , A61K35/30 , G01N33/50 , C12N5/0793
CPC分类号： C12N5/0621 , A61K35/30 , A61K35/44 , A61L27/00 , A61L27/383 , A61L27/3895 , C12N5/0619 , C12N2501/115 , C12N2501/155 , C12N2501/41 , C12N2501/50 , C12N2501/999 , C12N2506/45 , C12N2533/52 , C12Q1/02 , G01N33/5014 , G01N33/5058
摘要： The present invention provides a method for producing retinal cells or a retinal tissue, comprising the following steps (1)-(3): (1) a first step of culturing human pluripotent stem cells in the absence of feeder cells and in a medium comprising a factor for maintaining undifferentiated state, (2) a second step of culturing the pluripotent stem cells obtained in the first step in suspension in the presence of a Sonic hedgehog signal transduction pathway activating substance to form a cell aggregate, and (3) a third step of culturing the aggregate obtained in the second step in suspension in the presence of a 1) a BMP signal transduction pathway activating substance to obtain an aggregate containing retinal cells or a retinal tissue.
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公开(公告)号：US20190169570A1
公开(公告)日：2019-06-06
申请号：US16267948
申请日：2019-02-05
发明人： Atsushi KUWAHARA , Yoshiki SASAI
摘要： The present invention provides a method for producing a cell aggregate containing a ciliary marginal zone-like structure, including a step of culturing a cell aggregate containing a retinal tissue in which Chx10 positive cells are present in a proportion of 20% or more and 100% or less of the tissue in a serum-free medium or serum-containing medium each containing a substance acting on the Wnt signal pathway and a substance inhibiting the FGF signal pathway for only a period before the appearance of a RPE65 gene expressing cell, followed by culturing the “cell aggregate in which a RPE65 gene-expressing cell does not appear” thus obtained in a serum-free medium or serum-containing medium each free of a substance acting on the Wnt signal pathway and so on. According to the production method of the present invention, a ciliary marginal zone-like structure can be produced with high efficiency.
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公开(公告)号：US20170319748A1
公开(公告)日：2017-11-09
申请号：US15112187
申请日：2014-10-16
IPC分类号： A61L27/38 , A61K35/545 , C12N5/079
CPC分类号： A61L27/3878 , A61K35/30 , A61K35/545 , A61L27/383 , A61L27/3834 , A61L27/3895 , A61L2400/06 , A61L2430/16 , C12N5/0621 , C12N2501/10 , C12N2501/11 , C12N2501/115 , C12N2501/155 , C12N2501/385 , C12N2501/41 , C12N2501/415 , C12N2501/727 , C12N2501/999 , C12N2506/02
摘要： The invention provides a method for producing a ciliary marginal zone stem cell induced to differentiate from a pluripotent stem cell, including either the following step (1) or step (2), or both of these steps: (1) a step of floating culturing cells obtained from a cell aggregate containing a ciliary marginal zone-like structure induced to differentiate from pluripotent stem cells, thereby obtaining a retinosphere; and (2) a step of collecting stage specific embryonic antigen-1 positive cells from cells obtained from a cell aggregate containing a ciliary marginal zone-like structure induced to differentiate from pluripotent stem cells.
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公开(公告)号：US20160376554A1
公开(公告)日：2016-12-29
申请号：US15103162
申请日：2014-10-16
发明人： Atsushi KUWAHARA , Yoshiki SASAI
CPC分类号： C12N5/0621 , A61K9/0051 , A61K35/30 , A61L27/3641 , A61L27/3687 , A61L27/3804 , A61L2300/64 , A61L2430/16 , C12N2500/90 , C12N2501/119 , C12N2501/155 , C12N2501/385 , C12N2501/415 , C12N2501/727 , C12N2501/73 , C12N2506/02 , C12N2506/45 , C12N2513/00 , G01N33/5008 , G01N33/5014
摘要： The present invention provides a method for producing a cell aggregate containing a ciliary marginal zone-like structure, including a step of culturing a cell aggregate containing a retinal tissue in which Chx10 positive cells are present in a proportion of 20% or more and 100% or less of the tissue in a serum-free medium or serum-containing medium each containing a substance acting on the Wnt signal pathway and a substance inhibiting the FGF signal pathway for only a period before the appearance of a RPE65 gene expressing cell, followed by culturing the “cell aggregate in which a RPE65 gene-expressing cell does not appear” thus obtained in a serum-free medium or serum-containing medium each free of a substance acting on the Wnt signal pathway and so on. According to the production method of the present invention, a ciliary marginal zone-like structure can be produced with high efficiency.
摘要翻译： 本发明提供了一种含有睫状边缘区域结构的细胞聚集体的制造方法，其特征在于，包括培养包含视网膜组织的细胞集合体的步骤，其中Chx10阳性细胞以20％以上且100％以上的比例存在， 或更少的无血清培养基或含血清的培养基中含有作用于Wnt信号通路的物质的组织，以及抑制FGF信号通路的物质仅在RPE65基因表达细胞出现之前的一段时间后， 在不含有作用于Wnt信号通路的物质的无血清培养基或含血清培养基中培养“RPE65基因表达细胞未出现的细胞集合体”等。 根据本发明的制造方法，可以高效率地制造睫状边缘区域状结构体。
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公开(公告)号：US20190112575A1
公开(公告)日：2019-04-18
申请号：US16095092
申请日：2017-04-24
IPC分类号： C12N5/0793 , C12N5/074
摘要： A cell population comprising Corin- and/or Lrtm1-positive cells was produced by the following steps (1) and (2), from which Corin positive and/or Lrtm1 positive cells are collected using a substance that binds to Corin and/or a substance that binds to Lrtm1, and dopaminergic neuron progenitor cells are produced by performing suspension culture of the Corin positive and/or Lrtm1 positive cells in a culture solution containing one or more nutritional factors: (1) a step of performing adhesion culture of pluripotent stem cells in a medium for maintaining undifferentiated state containing a Sonic hedgehog (SHH) signal stimulant, and an undifferentiated state-maintaining factor in the absence of feeder cells but in the presence of an extracellular matrix, and (2) a step of culturing the cell population obtained in the step (1) in a culture solution containing one or more differentiation-inducing factors.
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