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    • 7. 发明申请
    • DEFINED MEDIA FOR PLURIPOTENT STEM CELL CULTURE
    • 定义的细胞培养基
    • WO2005065354A2
    • 2005-07-21
    • PCT/US2004/043858
    • 2004-12-31
    • THE BURNHAM INSTITUTEPARSONS, Xuejun, HuangSNYDER, Evan, Y.
    • PARSONS, Xuejun, HuangSNYDER, Evan, Y.
    • A01N63/00A61K35/12C12N5/00C12N5/02C12N5/0735C12N5/074C12N5/10G01N33/50
    • C12N5/0606A61K35/12C12N5/0607C12N2500/38C12N2500/44C12N2500/90C12N2500/98C12N2501/115C12N2501/33C12N2502/13C12N2503/00C12N2533/52C12N2533/54G01N33/5073
    • Stem cells, including mammalian, and particularly primate primordial stem cells (pPSCs) such as human embryonic stem cells (hESCs), hold great promise for restoring cell, tissue, and organ function. However, cultivation of stem cells, particularly undifferentiated hESCs, in serum-free, feeder-free, and conditioned-medium free conditions remain crucial for large-scale, uniform production of pluripotent cells for cell-based therapies, as well as for controlling conditions for efficiently directing their lineage-specific differentiation. This instant invention is based on the discovery of the formulation of minimal essential components necessary for maintaining the long-term growth of pPSCs, particularly undifferentiated hESCs. Basic fibroblast growth factor (bFGF), insulin, ascorbic acid, and laminin were identified to be both sufficient and necessary for maintaining hESCs in a healthy self-renewing undifferentiated state capable of both prolonged propagation and then directed differentiation. Having discerned these minimal molecular requirements, conditions that would permit the substitution of poorly-characterized and unspecified biological additives and Substrates were derived and optimized with entirely defined constituents, providing a “biologics”-free( i.e. , animal-, feeder-, serum-, and conditioned-medium-free) system for the efficient long-term cultivation of pPSCs, particularly pluripotent hESCs. Such culture systems allow the derivation and large-scale production of stem cells such as pPSCs, particularly pluripotent hESCs, in optimal yet well-defined biologics-free culture conditions from which they can be efficiently directed towards a lineage-specific differentiated fate in vitro , and thus are important, for instance, in connection with clinical applications based on stem cell therapy and in drug discovery processes.
    • 干细胞,包括哺乳动物,特别是灵长类动物原始干细胞(pPSCs)如人类胚胎干细胞(hESCs),对恢复细胞,组织和器官功能具有重要的前景。 然而,干细胞,特别是未分化的hESCs的培养在无血清,无饲养细胞和无条件培养基的条件下对于用于基于细胞的疗法的多能细胞的大规模,均匀生产以及用于控制条件 以有效地指导其谱系特异性分化。 本发明基于发现维持pPSC,特别是未分化hESC的长期生长所必需的最小必需成分的配方。 碱性成纤维细胞生长因子(bFGF),胰岛素,抗坏血酸和层粘连蛋白被认为既足够又是必需的,以维持hESCs处于能够延长繁殖,然后进行定向分化的健康自我更新未分化状态。 鉴于这些最小的分子要求,可以使用完全限定的成分衍生和优化允许代表不良表征和未指定的生物添加剂和底物的条件,提供“生物制剂”(即动物,饲料,血清 - ,无条件培养基)系统,用于高效长期培养pPSCs,特别是多能性hESC。 这样的培养系统允许干细胞的衍生和大规模生产,例如pPSC,特别是多能hESC,在最佳且明确定义的不含生物制剂的培养条件中,从而可以有效地将其引导到体外特有的分化命运, 因此例如与基于干细胞治疗和药物发现过程的临床应用相关是重要的。