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    Method for culturing skeletal muscle for tissue engineering
    2.
    发明授权
    Method for culturing skeletal muscle for tissue engineering 有权
    组织工程骨骼肌培养方法

    公开(公告)号：US09163216B1

    公开(公告)日：2015-10-20

    申请号：US12765399

    申请日：2010-04-22

    申请人： James J. Hickman ,  Mainak Das ,  John W. Rumsey

    发明人： James J. Hickman ,  Mainak Das ,  John W. Rumsey

    IPC分类号： C12N5/077 ,  C12N5/0793

    CPC分类号： C12N5/0652 ,  C12N5/0619 ,  C12N5/0658 ,  C12N2500/05 ,  C12N2500/46 ,  C12N2500/90 ,  C12N2500/99 ,  C12N2501/40 ,  C12N2502/1335 ,  C12N2533/30

    摘要： The invention provides a nutrient medium composition and associated methods for lengthening the useful life of a culture of muscle cells. Disclosed is a method of culturing mammalian muscle cells, including preparing one or more carriers coated with a covalently bonded monolayer of trimethoxy-silylpropyl-diethylenetriamine (DETA); verifying DETA monolayer formation by one or more associated optical parameters; suspending isolated fetal rat skeletal muscle cells in serum-free medium according to medium composition 1; plating the suspended cells onto the prepared carriers at a predetermined density; leaving the carriers undisturbed for cells to adhere to the DETA monolayer; covering the carriers with a mixture of medium 1 and medium 2; and incubating. A cell nutrient medium composition includes Neurobasal, an antibiotic-antimycotic composition, cholesterol, human TNF-alpha, PDGF BB, vasoactive intestinal peptides, insulin-like growth factor 1, NAP, r-Apolipoprotein E2, purified mouse Laminin, beta amyloid, human tenascin-C protein, rr-Sonic hedgehog Shh N-terminal, and rr-Agrin C terminal.

    摘要翻译： 本发明提供了一种营养培养基组合物和相关方法，用于延长肌肉细胞培养物的使用寿命。 公开了一种培养哺乳动物肌肉细胞的方法，包括制备一种或多种涂覆有共价键合的三甲氧基 - 甲硅烷基丙基 - 二亚乙基三胺（DETA）单层的载体; 通过一个或多个相关联的光学参数验证DETA单层形成; 根据培养基组成1，在无血清培养基中悬浮分离的胎鼠大鼠骨骼肌细胞; 以预定的密度将悬浮细胞电镀到制备的载体上; 使载体不受干扰以使细胞粘附到DETA单层上; 用介质1和介质2的混合物覆盖载体; 并孵化。 细胞营养培养基组合物包括Neurobasal，抗生素 - 抗真菌组合物，胆固醇，人TNF-α，PDGF BB，血管活性肠肽，胰岛素样生长因子1，NAP，r-载脂蛋白E2，纯化的小鼠层粘连蛋白，β淀粉样蛋白，人 腱生蛋白-C蛋白，rr-Sonic刺猬Shh N末端和rr-Agrin C端。