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    • 75. 发明申请
    • NOVEL PROCESS
    • 新进程
    • WO2011047900A2
    • 2011-04-28
    • PCT/EP2010/062103
    • 2010-08-19
    • GLAXOSMITHKLINE BIOLOGICALS S.A.GERARD, Jacques, Dominique, MarieKNOTT, Isabelle, Solange, Lucie
    • GERARD, Jacques, Dominique, MarieKNOTT, Isabelle, Solange, Lucie
    • C12M47/02C12M25/16
    • The present invention relates to a process for detaching cells of an adherent cell type from particles of carrier which comprises the steps of: (a) providing in a vessel said cells adhered to particles of carrier in the presence of a growth medium, said vessel being characterised in that it comprises two internal zones, A and B, separated by a sieve, the sieve having an aperture size such that it is pervious to the cells if detached from the particles of carrier but substantially impervious to the particles of carrier, whereby the particles of carrier having cells adhered thereto are located within zone A but not zone B and zone B contains a drain opening under control of a valve adapted to permit vacation of liquid from zone A under an influence comprising gravity and/or pressure; (b) optionally washing the particles of carrier having cells adhered thereto by vacating liquid from the vessel via the drain opening in zone B under an influence comprising gravity and/or pressure and replenishing the vessel with another wash liquid and optionally repeating this step one or more times; (c) replacing the liquid in the vessel with a detachment buffer by vacating liquid from the vessel via the drain opening in zone B under an influence comprising gravity and/or pressure and replenishing the vessel with the detachment buffer; (c) detaching the cells from the particles of carrier by a process comprising an enzymatic detachment step performed in a detachment buffer.
    • 本发明涉及从载体颗粒中分离粘附细胞类型的细胞的方法,其包括以下步骤:(a)在生长培养基存在下,在容器中提供附着于载体颗粒的细胞,所述容器为 其特征在于它包括由筛子分隔的两个内部区域A和B,所述筛子具有孔径尺寸,使得如果从载体颗粒分离而基本上不透过载体颗粒,其可以透过细胞,由此 具有附着细胞的细胞的颗粒位于区域A内,而不是区域B,而区域B在适于在包含重力和/或压力的影响下允许从区域A中排出液体的阀控制下包含排水口; (b)任选地在包含重力和/或压力的影响下通过区域B中的排水开口从容器中排出液体来洗涤附着细胞的载体颗粒,并用另一种洗涤液补充该容器,并任选重复该步骤一或 更多次 (c)通过在包含重力和/或压力的影响下通过区域B中的排水开口从容器中排出液体,用分离缓冲液代替容器中的液体,并用分离缓冲液补充容器; (c)通过包括在脱离缓冲液中进行的酶解脱离步骤的方法从载体颗粒中分离细胞。