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    • 1. 发明申请
    • PANCREATIC ISLET SEPARATION METHOD, AND PROTECTIVE SOLUTION FOR PROTECTING PANCREATIC ISLET TISSUE
    • 胰岛分离方法和保护胰腺组织的保护性解决方案
    • US20130177986A1
    • 2013-07-11
    • US13808390
    • 2011-07-13
    • Masahiro TanemuraYoshiki SawaAkira MyouriToshinori ItoMasaki MoriYuichiro Doki
    • Masahiro TanemuraYoshiki SawaAkira MyouriToshinori ItoMasaki MoriYuichiro Doki
    • C12N5/071
    • C12N5/0676C12N2501/734
    • The disclosed islet isolation method comprises: an injection step of injecting a preservation solution into the pancreatic duct of an excised pancreas; a preservation step of immersing the pancreas into an immersion fluid for preservation; a digestion step of breaking down the pancreas to provide pancreatic tissue; and a purification step of immersing the pancreatic tissue in a purification solution to provide islets. The digestion step consists of: an enzyme injection step of injecting an enzyme solution containing a digestion enzyme into the pancreas; a digestion initiation step of activating the digestion enzyme; a digestion termination step of inactivating the digestion enzyme; and a collection step of collecting the broken-down pancreatic tissue. The islet isolation method is characterized in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation step, the neutrophil elastase inhibitor is present inside the pancreas at the time point of starting the digestion initiation step. By using the above method and a protective solution which can be used in the method, islets having a size and shape suitable for transplantation can be obtained in high yields.
    • 公开的胰岛分离方法包括:将保存液注入切除胰腺胰管的注射步骤; 将胰脏浸入用于保存的浸液中的保存步骤; 消化胰腺胰腺组织消化步骤; 以及将胰组织浸入净化溶液中以提供胰岛的纯化步骤。 消化步骤包括:将含有消化酶的酶溶液注入胰腺的酶注射步骤; 活化消化酶的消化引发步骤; 消化终止步骤,使消化酶失活; 以及收集分解的胰组织的收集步骤。 胰岛分离方法的特征在于,通过在消化引发步骤之前向系统中加入嗜中性粒细胞弹性蛋白酶抑制剂,在开始消化起始步骤的时间点,存在嗜中性粒细胞弹性蛋白酶抑制剂在胰腺内。 通过使用上述方法和可用于该方法的保护溶液,可以高产率获得具有适合于移植的尺寸和形状的胰岛。
    • 2. 发明授权
    • Pancreatic islet separation method, and protective solution for protecting pancreatic islet tissue
    • 胰岛分离法,保护胰岛组织保护液
    • US08759099B2
    • 2014-06-24
    • US13808390
    • 2011-07-13
    • Masahiro TanemuraYoshiki SawaAkira MyouiToshinori ItoMasaki MoriYuichiro Doki
    • Masahiro TanemuraYoshiki SawaAkira MyouiToshinori ItoMasaki MoriYuichiro Doki
    • C12N5/00
    • C12N5/0676C12N2501/734
    • The disclosed islet isolation method comprises: an injection step of injecting a preservation solution into the pancreatic duct of an excised pancreas; a preservation step of immersing the pancreas into an immersion fluid for preservation; a digestion step of breaking down the pancreas to provide pancreatic tissue; and a purification step of immersing the pancreatic tissue in a purification solution to provide islets. The digestion step consists of: an enzyme injection step of injecting an enzyme solution containing a digestion enzyme into the pancreas; a digestion initiation step of activating the digestion enzyme; a digestion termination step of inactivating the digestion enzyme; and a collection step of collecting the broken-down pancreatic tissue. The islet isolation method is characterized in that, by adding a neutrophil elastase inhibitor to the system before the digestion initiation step, the neutrophil elastase inhibitor is present inside the pancreas at the time point of starting the digestion initiation step. By using the above method and a protective solution which can be used in the method, islets having a size and shape suitable for transplantation can be obtained in high yields.
    • 公开的胰岛分离方法包括:将保存液注入切除胰腺胰管的注射步骤; 将胰脏浸入用于保存的浸液中的保存步骤; 消化胰腺胰腺组织消化步骤; 以及将胰组织浸入净化溶液中以提供胰岛的纯化步骤。 消化步骤包括:将含有消化酶的酶溶液注入胰腺的酶注射步骤; 活化消化酶的消化引发步骤; 消化终止步骤,使消化酶失活; 以及收集分解的胰组织的收集步骤。 胰岛分离方法的特征在于,通过在消化引发步骤之前向系统中加入嗜中性粒细胞弹性蛋白酶抑制剂,在开始消化起始步骤的时间点,存在嗜中性粒细胞弹性蛋白酶抑制剂在胰腺内。 通过使用上述方法和可用于该方法的保护溶液,可以高产率获得具有适合于移植的尺寸和形状的胰岛。
    • 7. 发明申请
    • Method of producing three-dimensional tissue and method of producing extracellular matrix used in the same
    • 制备三维组织的方法及其制备方法
    • US20070207540A1
    • 2007-09-06
    • US11514468
    • 2006-09-01
    • Mitsuru AkashiYoshiki SawaMichiya MatsusakiYoshio Nakahara
    • Mitsuru AkashiYoshiki SawaMichiya MatsusakiYoshio Nakahara
    • C12N5/06
    • C12N5/0068C12N5/0062C12N2533/52C12N2533/54Y10S977/902
    • The present invention provides a novel method of producing a three-dimensional tissue by which cell lamination can be carried out easily. According to the method, a three-dimensional tissue in which cell layers are laminated with an extracellular matrix intervening between each pair of the adjacent cell layers is produced by: (A) forming a cell layer on a substrate; (B) bringing the cell layer formed on the substrate into contact with a solution containing a first substance and a solution containing a second substance alternately, thus forming, on the cell layer, an extracellular matrix in which the first substance and the second substance are laminated alternately; and (C) culturing a cell on the extracellular matrix to form a further cell layer. In the present invention, a combination of the first substance and the second substance is (a) a combination of a protein or polymer having an RGD sequence and a protein or polymer that interacts with the protein or polymer having the RGD sequence or (b) a combination of a protein or polymer that is positively charged and a protein or polymer that is negatively charged.
    • 本发明提供一种可以容易地进行细胞层压的三维组织的制造方法。 根据该方法,通过以下方法制造细胞层与介于每对相邻细胞层之间的细胞外基质层叠的三维组织:(A)在基材上形成细胞层; (B)使形成在基板上的细胞层与含有第一物质的溶液和含有第二物质的溶液交替接触,从而在细胞层上形成第一物质和第二物质为细胞外基质 交替叠层; 和(C)在细胞外基质上培养细胞以形成另外的细胞层。 在本发明中,第一物质和第二物质的组合是(a)具有RGD序列的蛋白质或聚合物和与具有RGD序列的蛋白质或聚合物相互作用的蛋白质或聚合物的组合,或(b) 带正电荷的蛋白质或聚合物和带负电荷的蛋白质或聚合物的组合。