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1. 发明申请

US20130323839A1 Culture Substrate and Culture Sheet 审中-公开
标题翻译：文化底物和文化表
公开(公告)号：US20130323839A1
公开(公告)日：2013-12-05
申请号：US13996074
申请日：2010-12-22
申请人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda , Taku Saito , Naoshi Itabashi , Jiro Yamamoto
发明人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda , Taku Saito , Naoshi Itabashi , Jiro Yamamoto
IPC分类号： G01N33/50
CPC分类号： G01N33/5008 , C12M25/00 , C12M25/06
摘要： Provided is a culture sheet which enables a technique for forming a three-dimensional tissue having uniform diameter without applying any chemical on the surface of a culture substrate. On the culture sheet (150) of the culture substrate, a plurality of holes (152) are formed and nanopillars (153), which are capable of controlling the adhesiveness and migration ability of cells, are formed on the bottom surface of each hole (152), said bottom face serving as a culture surface. The culture surface of each hole (151) is provided with a partition wall (152) and the internal nanopillars (153) are formed in the vicinity of the center of the hole (151). Owing to this configuration, the interaction among the disseminated cells can be restricted so that uniformly sized three-dimensional structures of the cells can be formed.
摘要翻译：提供一种培养片，其能够形成具有均匀直径的三维组织的技术，而不在培养基材的表面上施加任何化学物质。在培养基板的培养板（150）上形成有多个孔（152），并且能够控制细胞的粘附性和迁移能力的纳米柱（153）形成在各孔的底面 152），所述底面用作培养表面。每个孔（151）的培养表面设置有分隔壁（152），并且内孔纳米柱（153）形成在孔（151）的中心附近。由于这种构造，可以限制扩散细胞之间的相互作用，从而可以形成均匀尺寸的细胞的三维结构。

2. 发明申请

US20120100612A1 CULTURE SUBSTRATE, CULTURE SHEET, AND CELL CULTURE METHOD 有权
标题翻译：文化底物，文化表和细胞培养方法
公开(公告)号：US20120100612A1
公开(公告)日：2012-04-26
申请号：US13379416
申请日：2010-06-22
申请人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda , Taku Saito
发明人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda , Taku Saito
IPC分类号： C12N5/0735 , C12N5/071 , C12M3/00
CPC分类号： C12N5/0062 , C12M21/08 , C12M23/12 , C12M25/06 , C12N2535/10
摘要： Disclosed is a culture sheet which enables technology in which three-dimensional tissues with uniform diameter are formed without applying chemicals to the surface of a culture substrate. A plurality of holes are formed on the culture sheet of the substrate, and nanopillars capable of controlling the adhesiveness or migration of a cell are formed on a culture surface that serves as the bottom surface of each of the holes. The culture surface of each of the holes having a structure in which a partition wall is provided, wherein, by forming the internal nanopillars in the vicinity of the center of each of the holes, the interaction of the disseminated cells can be limited to uniform the size of the three-dimensional structures of the cells to be formed.
摘要翻译：公开了一种能够形成均匀直径的三维组织而不向培养基材表面施加化学物质的培养片。在基板的培养片上形成多个孔，在作为各孔的底面的培养面上形成能够控制细胞的粘附性或迁移的纳米柱。每个孔的培养面具有其中设置隔壁的结构，其中，通过在每个孔的中心附近形成内部纳米柱，可以将扩散细胞的相互作用限制为均匀的要形成的细胞的三维结构的尺寸。

3. 发明授权

US09394514B2 Culture substrate, culture sheet, and cell culture method 有权
标题翻译：培养基质，培养片和细胞培养法
公开(公告)号：US09394514B2
公开(公告)日：2016-07-19
申请号：US13379416
申请日：2010-06-22
申请人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda , Taku Saito
发明人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda , Taku Saito
IPC分类号： C12M3/00 , C12N5/00 , C12M1/32 , C12M1/12
CPC分类号： C12N5/0062 , C12M21/08 , C12M23/12 , C12M25/06 , C12N2535/10
摘要： Disclosed is a culture sheet which enables technology in which three-dimensional tissues with uniform diameter are formed without applying chemicals to the surface of a culture substrate. A plurality of holes are formed on the culture sheet of the substrate, and nanopillars capable of controlling the adhesiveness or migration of a cell are formed on a culture surface that serves as the bottom surface of each of the holes. The culture surface of each of the holes having a structure in which a partition wall is provided, wherein, by forming the internal nanopillars in the vicinity of the center of each of the holes, the interaction of the disseminated cells can be limited to uniform the size of the three-dimensional structures of the cells to be formed.
摘要翻译：公开了一种能够形成均匀直径的三维组织而不向培养基材表面施加化学物质的培养片。在基板的培养片上形成多个孔，在作为各孔的底面的培养面上形成能够控制细胞的粘附性或迁移的纳米柱。每个孔的培养面具有其中设置隔壁的结构，其中，通过在每个孔的中心附近形成内部纳米柱，可以将扩散细胞的相互作用限制为均匀的要形成的细胞的三维结构的尺寸。

4. 发明申请

US20140162351A1 CULTURING SHEET, CULTURING EQUIPMENT MATERIAL AND MANUFACTURING METHOD 审中-公开
标题翻译：文化用品，文化设备材料及制造方法
公开(公告)号：US20140162351A1
公开(公告)日：2014-06-12
申请号：US14131767
申请日：2011-08-29
申请人： Jiro Yamamoto , Naoshi Itabashi , Taku Saito , Akiko Hisada , Ryosuke Takahashi , Hiroshi Sonoda
发明人： Jiro Yamamoto , Naoshi Itabashi , Taku Saito , Akiko Hisada , Ryosuke Takahashi , Hiroshi Sonoda
IPC分类号： C12N5/071 , B29C59/02 , C12N5/04
CPC分类号： C12N5/0602 , B29C59/022 , C12M23/12 , C12M25/00 , C12N5/04
摘要： An operating efficiency of an observer is considerably restricted since it is not known at which position a culturing cell is disposed among a great number of pieces of holes of a culturing sheet. The culturing sheet is configured by a partitioning wall, a hole isolated by the partitioning wall, a local culturing region formed with plural local culturing region pillars a height of which is lower than that of the partitioning wall at a portion of a bottom face, and identification mark pillars formed at an identification mark region which differs from the culturing region at the bottom face of the hole. An identification mark is prevented from being unable to be optically recognized by adhering a spheroid to the identification mark region by making a diameter and a height of the identification mark pillar smaller than a diameter and a height of the local culturing pillar.
摘要翻译：观察者的操作效率受到相当的限制，因为在培养板的大量孔中不设置培养单元的位置。培养片由分隔壁，由分隔壁隔离的空穴构成，局部培养区形成有多个局部培养区柱，其高度低于底面部分的分隔壁，识别标记柱形成在与孔底面的培养区域不同的识别标记区域。通过使识别标记柱的直径和高度小于局部培养柱的直径和高度，通过将球体粘附到识别标记区域来防止识别标记不被光学识别。

5. 发明申请

US20110269232A1 METHOD FOR CULTURING ANIMAL HEPATOCYTE 有权
标题翻译：培养动物肝细胞的方法
公开(公告)号：US20110269232A1
公开(公告)日：2011-11-03
申请号：US13143569
申请日：2009-01-08
申请人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda
发明人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda
IPC分类号： C12N5/071 , B82Y99/00
CPC分类号： C12N5/0671 , C12N2500/25 , C12N2501/39 , C12N2533/54
摘要： Provided are a technique for easily forming a spheroid by three-dimensionally culturing hepatocytes, and a technique for forming a spheroid having a higher expression level of a transporter MRP2 playing a role of biliary excretion than that of a conventional method. In order to solve the above-described problems, the present inventors have found out a condition under which hepatocytes easily form the spheroid on a nanopillar sheet. More specifically, this is related to a concentration of Type I collagen coated onto the NP sheet. Also, they have found out a condition under which an expression level of a gene related to the excretion of the formed spheroid is improved. More specifically, after the spheroid is previously formed, a biological matrix is overlayered thereon.
摘要翻译：提供通过三维培养肝细胞容易地形成球状体的技术，以及形成具有比常规方法具有胆汁排泄作用的转运蛋白MRP2更高表达水平的球状体的技术。为了解决上述问题，本发明人发现肝细胞容易在纳米柱片材上形成球状体的条件。更具体地说，这涉及涂覆在NP片上的I型胶原蛋白的浓度。此外，他们发现了与形成的球体的排泄相关的基因的表达水平提高的条件。更具体地说，在预先形成球体之后，将生物体叠加在其上。

6. 发明授权

US08530237B2 Method for culturing animal hepatocyte 有权
标题翻译：培养动物肝细胞的方法
公开(公告)号：US08530237B2
公开(公告)日：2013-09-10
申请号：US13143569
申请日：2009-01-08
申请人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda
发明人： Ryosuke Takahashi , Akiko Hisada , Hiroshi Sonoda
IPC分类号： C12N5/02 , A01N63/00
CPC分类号： C12N5/0671 , C12N2500/25 , C12N2501/39 , C12N2533/54
摘要： Provided are a technique for easily forming a spheroid by three-dimensionally culturing hepatocytes, and a technique for forming a spheroid having a higher expression level of a transporter MRP2 playing a role of biliary excretion than that of a conventional method. In order to solve the above-described problems, the present inventors have found out a condition under which hepatocytes easily form the spheroid on a nanopillar sheet. More specifically, this is related to a concentration of Type I collagen coated onto the NP sheet. Also, they have found out a condition under which an expression level of a gene related to the excretion of the formed spheroid is improved. More specifically, after the spheroid is previously formed, a biological matrix is overlayered thereon.
摘要翻译：提供通过三维培养肝细胞容易地形成球状体的技术，以及形成具有比常规方法具有胆汁排泄作用的转运蛋白MRP2更高表达水平的球状体的技术。为了解决上述问题，本发明人发现肝细胞容易在纳米柱片材上形成球状体的条件。更具体地说，这涉及涂覆在NP片上的I型胶原蛋白的浓度。此外，他们发现了与形成的球体的排泄相关的基因的表达水平提高的条件。更具体地说，在预先形成球体之后，将生物体叠加在其上。

7. 发明授权

US08043852B2 Monoclonal antibody specific to dentin-derived heparan sulfate 有权
标题翻译：牙本质硫酸乙酰肝素特异性单克隆抗体
公开(公告)号：US08043852B2
公开(公告)日：2011-10-25
申请号：US12222889
申请日：2008-08-19
申请人： Akiko Hisada , Ryosuke Takahashi , Hiroko Hanzawa
发明人： Akiko Hisada , Ryosuke Takahashi , Hiroko Hanzawa
IPC分类号： C12N5/12 , C12N5/00 , C07K16/00 , C07K16/18 , G01N33/53 , G01N33/577
CPC分类号： C07K16/18 , G01N2800/18
摘要： The present invention provides a monoclonal antibody displaying excellent specificity against heparan sulfate saccharide chains for the analysis of heparan sulfate saccharide chains specific to dentin. The invention also provides a method of evaluating reproductive dentin using the monoclonal antibody. The anti-heparan sulfate monoclonal antibody reacts against dentin-derived heparan sulfate and in particular the anti-heparan sulfate monoclonal antibody reacts strongly and specifically with uncalcified predentin regions. In the method of evaluating dentin, the antibody is reacted against an isolated dentin-derived sample and the reaction is used in order to evaluate the development of dentin.
摘要翻译：本发明提供了对硫酸乙酰肝糖链具有优异特异性的单克隆抗体，用于分析牙本质特异性的硫酸乙酰肝糖链。本发明还提供了使用单克隆抗体评估生殖牙本质的方法。抗硫酸乙酰肝素单克隆抗体与牙本质来源的硫酸乙酰肝素反应，特别是抗硫酸乙酰肝素单克隆抗体强烈而特异地与未煅烧的前体蛋白区域反应。在评估牙本质的方法中，抗体与分离的牙本质衍生的样品反应，并且使用反应来评估牙本质的发育。

8. 发明申请

US20090081809A1 Monoclonal antibody specific to dentin-derived heparan sulfate 有权
标题翻译：牙本质硫酸乙酰肝素特异性单克隆抗体
公开(公告)号：US20090081809A1
公开(公告)日：2009-03-26
申请号：US12222889
申请日：2008-08-19
申请人： Akiko Hisada , Ryossuke Takahashi , Hiroko Hanzawa
发明人： Akiko Hisada , Ryossuke Takahashi , Hiroko Hanzawa
IPC分类号： G01N33/566 , C07K16/18 , C12N5/06
CPC分类号： C07K16/18 , G01N2800/18
摘要： The present invention provides a monoclonal antibody displaying excellent specificity against heparan sulfate saccharide chains for the analysis of heparan sulfate saccharide chains specific to dentin. The invention also provides a method of evaluating reproductive dentin using the monoclonal antibody. The anti-heparan sulfate monoclonal antibody reacts against dentin-derived heparan sulfate and in particular the anti-heparan sulfate monoclonal antibody reacts strongly and specifically with uncalcified predentin regions. In the method of evaluating dentin, the antibody is reacted against an isolated dentin-derived sample and the reaction is used in order to evaluate the development of dentin.
摘要翻译：本发明提供了对硫酸乙酰肝糖链具有优异特异性的单克隆抗体，用于分析牙本质特异性的硫酸乙酰肝糖链。本发明还提供了使用单克隆抗体评估生殖牙本质的方法。抗硫酸乙酰肝素单克隆抗体与牙本质来源的硫酸乙酰肝素反应，特别是抗硫酸乙酰肝素单克隆抗体强烈而特异地与未煅烧的前体蛋白区域反应。在评估牙本质的方法中，抗体与分离的牙本质衍生的样品反应，并且使用反应来评估牙本质的发育。

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