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    Method for enhancing activity of laminin fragments as cell culture matrix
    2.
    发明授权
    Method for enhancing activity of laminin fragments as cell culture matrix 有权

    公开(公告)号：US10428311B2

    公开(公告)日：2019-10-01

    申请号：US15325743

    申请日：2015-07-15

    Applicant: OSAKA UNIVERSITY

    Inventor： Kiyotoshi Sekiguchi ,  Ko Tsutsui

    IPC: C12N5/074 ,  C12N15/00 ,  C12N15/09

    Abstract: The present invention provides a novel technique in a cell culture method using a cell culture vessel coated with a laminin fragment, which novel technique achieves cell culture as in the case of using a recommended coating concentration even when the coating concentration is lower than the recommended coating concentration. The present invention relates to a method for enhancing an activity for mammalian cultured cells of a laminin fragment or a variant thereof each having integrin binding activity.

    Cell culture vessel coated with laminin fragment in dry state
    6.
    发明授权
    Cell culture vessel coated with laminin fragment in dry state 有权

    公开(公告)号：US10287541B2

    公开(公告)日：2019-05-14

    申请号：US14897298

    申请日：2014-05-09

    Applicant: OSAKA UNIVERSITY

    Inventor： Kiyotoshi Sekiguchi ,  Ko Tsutsui

    IPC: C12M1/00 ,  C12M1/12 ,  C12N5/00 ,  C12N9/10 ,  C07K14/78 ,  C07K14/79 ,  C07K14/435 ,  C07K14/765

    Abstract: Provided is a cell culture vessel characterized in that a surface to be in contact with cells is coated with a laminin fragment having integrin α6β1 binding activity or a modified form thereof in a dry state, the laminin fragment being derived from at least one kind selected from laminin α5β1γ1 and laminin α5β2γ1, the cell culture vessel being any of the following: (1) a cell culture vessel of which a surface to be in contact with cells is coated only with a laminin fragment having integrin α6β1 binding activity or a modified form thereof in a dry state; (2) a cell culture vessel of which a surface to be in contact with cells is coated with a laminin fragment having integrin α6β1 binding activity or a modified form thereof in combination with a laminin fragment having no integrin α6β1 binding activity in a dry state; and (3) a cell culture vessel of which a surface to be in contact with cells is coated with a laminin fragment having integrin α6β1 binding activity or a modified form thereof in combination with a protein that is neither a laminin nor a laminin fragment, in a dry state.

    METHOD FOR CONTROLLING DIFFERENTIATION OF PLURIPOTENT STEM CELLS
    8.
    发明申请
    METHOD FOR CONTROLLING DIFFERENTIATION OF PLURIPOTENT STEM CELLS 审中-公开

    公开(公告)号：US20200010800A1

    公开(公告)日：2020-01-09

    申请号：US16482160

    申请日：2018-01-31

    Applicant: Osaka University

    Inventor： Kohji Nishida ,  Kiyotoshi Sekiguchi ,  Ryuhei Hayashi ,  Shun Shibata

    IPC: C12N5/079

    Abstract: The present invention relates to a method for controlling differentiation of pluripotent stem cells, which method comprises selecting a laminin or a fragment thereof based on binding affinity for the pluripotent stem cells and inducing differentiation of the pluripotent stem cells in the presence of the laminin or a fragment thereof. Here, the binding affinity for cells can be assessed by time-course observation of the survival rate and motility of the cells. According to the present invention, a cell population containing any desired proportion of differentiated cells can be produced from pluripotent stem cells in a simple manner. The cell population obtained by this production method is very useful for cell therapy-based treatment strategies for diseases.

    METHOD FOR INDUCING DOPAMINERGIC NEURON PROGENITOR CELLS
    10.
    发明申请
    METHOD FOR INDUCING DOPAMINERGIC NEURON PROGENITOR CELLS 审中-公开
    诱导多巴胺神经元原核细胞的方法

    公开(公告)号：US20160215260A1

    公开(公告)日：2016-07-28

    申请号：US14916696

    申请日：2014-09-04

    Applicant: KYOTO UNIVERSITY ,  OSAKA UNIVERSITY ,  EISAI R&D MANAGEMENT CO., LTD.

    Inventor： Jun Takahashi ,  Daisuke Doi ,  Bumpei Samata ,  Kiyotoshi Sekiguchi ,  Yuichi Ono

    IPC: C12N5/0797 ,  A61K35/30

    CPC classification number: C12N5/0623 ,  A61K35/30 ,  C12N5/0619 ,  C12N2501/119 ,  C12N2501/13 ,  C12N2501/15 ,  C12N2501/155 ,  C12N2501/41 ,  C12N2501/415 ,  C12N2501/727 ,  C12N2506/02 ,  C12N2506/03 ,  C12N2506/45 ,  C12N2533/52

    Abstract: The present invention provides a method for producing dopaminergic neuron progenitor cells from pluripotent stem cells, which method comprises the steps of: (i) performing adherent culture of pluripotent stem cells on an extracellular matrix in a medium containing a reagent(s) selected from the group consisting of BMP inhibitor, TGFβ inhibitor, SHH signal-stimulating agent, FGF8, and GSK3β inhibitor; (ii) collecting Corin- and/or Lrtm1-positive cells from the cells obtained in Step (i) using a substance which binds to Corin and/or a substance which binds to Lrtm1; and (iii) performing suspension culture of the cells obtained in Step (ii) in a medium containing a neurotrophic factor.

    Abstract translation: 本发明提供了一种用于从多能干细胞产生多巴胺能神经元祖细胞的方法，该方法包括以下步骤：（i）在含有选自以下的试剂的培养基中，在细胞外基质上进行多能干细胞的贴壁培养： 由BMP抑制剂，TGFβ抑制剂，SHH信号刺激剂，FGF8和GSK3β抑制剂组成的组; （ii）使用与Corin结合的物质和/或与Lrtm1结合的物质从步骤（i）中获得的细胞收集Corin和/或Lrtm1阳性细胞; 和（iii）在含有神经营养因子的培养基中进行步骤（ii）中获得的细胞的悬浮培养。

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