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    • 4. 发明申请
    • MEDIUM AND CULTURE OF EMBRYONIC STEM CELLS
    • 胚胎干细胞的培养和培养
    • US20120178161A1
    • 2012-07-12
    • US13427548
    • 2012-03-22
    • James A. ThomsonTenneille Ludwig
    • James A. ThomsonTenneille Ludwig
    • C12N5/071C12N5/0735
    • C12N5/0606C12N5/0696C12N2500/12C12N2500/20C12N2500/25C12N2500/32C12N2500/36C12N2500/38C12N2500/50C12N2500/90C12N2500/98C12N2501/115C12N2501/15C12N2501/845
    • Previous methods for culturing primate pluripotent stem cells have required either fibroblast feeder cells or a medium which was exposed to fibroblast feeder cells to maintain the stem cells in an undifferentiated state. It has now been found that high levels of fibroblast growth factor in a medium together with at least one of gamma aminobutyric acid, pipecolic acid, and lithium, enables pluripotent stem cells to remain undifferentiated indefinitely through multiple passages, even without feeder cells or conditioned medium. Without beta-mercaptoethanol, the medium improves cloning efficiency. Also, a matrix of human proteins can be used to culture the undifferentiated cells without exposing the cells to animal products. Further disclosed are new primate pluripotent cell lines made using the defined culture conditions, including the medium and the matrix. Such new cell lines will have never been exposed to animal cells, animal products, feeder cells or conditioned medium.
    • 培养灵长类多能干细胞的以前的方法需要成纤维细胞饲养细胞或暴露于成纤维细胞饲养细胞的培养基以维持干细胞处于未分化状态。 现在已经发现,培养基中高水平的成纤维细胞生长因子与γ-氨基丁酸,哌啶酸和锂中的至少一种一起使多能干细胞通过多次传代无限期地保持未分化,即使没有饲养细胞或条件培养基 。 没有β-巯基乙醇,培养基提高了克隆效率。 此外,可以使用人类蛋白质的基质来培养未分化细胞而不将细胞暴露于动物产品。 进一步公开的是使用定义的培养条件制备的新的灵长类动物多能细胞系,包括培养基和基质。 这样的新细胞系从未暴露于动物细胞,动物产品,饲养细胞或条件培养基。
    • 9. 发明授权
    • Primate embryonic stem cell line
    • 灵长类胚胎干细胞系
    • US07582479B2
    • 2009-09-01
    • US11036245
    • 2005-01-14
    • James A. Thomson
    • James A. Thomson
    • C12N15/02C12N15/04
    • C12N5/0605C12N5/0603C12N5/0606C12N2502/13C12N2506/02
    • A purified preparation of primate embryonic stem cells is disclosed. This preparation is characterized by the following cell surface markers: SSEA-1 (−); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); and alkaline phosphatase (+). In a particularly advantageous embodiment, the cells of the preparation are human embryonic stem cells, have normal karyotypes, and continue to proliferate in an undifferentiated state after continuous culture for eleven months. The embryonic stem cell lines also retain the ability, throughout the culture, to form trophoblast and to differentiate into all tissues derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). A method for isolating a primate embryonic stem cell line is also disclosed.
    • 公开了灵长类动物胚胎干细胞的纯化制剂。 该制剂的特征在于以下细胞表面标志物:SSEA-1( - ); SSEA-4(+); TRA-1-60(+); TRA-1-81(+); 和碱性磷酸酶(+)。 在一个特别有利的实施方案中,制剂的细胞是人胚胎干细胞,具有正常的核型,并且在连续培养11个月后,以未分化状态继续增殖。 胚胎干细胞系还保留了整个培养物中形成滋养细胞并分化成来自所有三个胚胎胚层(内胚层,中胚层和外胚层)的所有组织的能力。 还公开了分离灵长类动物胚胎干细胞系的方法。