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    CELL-BASED SHEAR STRESS SENSOR
    1.
    发明公开
    CELL-BASED SHEAR STRESS SENSOR 审中-实审

    公开(公告)号：EP4400592A2

    公开(公告)日：2024-07-17

    申请号：EP24178175.6

    申请日：2023-05-08

    申请人： Sartorius Stedim Biotech GmbH

    发明人： Akbari, Samin ,  Leroux, Ann-Cathrin ,  Kwon, Taehong ,  Zehe, Christoph ,  Pollard, David

    IPC分类号： C12N15/65

    CPC分类号： C12N15/65 ,  C12N15/85 ,  C12N2830/00220130101

    摘要： The present invention relates to a method of evaluating fluid shear stress acting on cells in real-time in a bioprocess and/or during the operation of a biotechnological device or system, comprising the steps of subjecting a cell to said bioprocess and/or to operation of said biotechnological device or system, wherein said cell comprises an isolated nucleic acid molecule or a nucleic acid vector comprising said isolated nucleic acid molecule, wherein said isolated nucleic acid molecule comprises a functional mammalian EGR-1 (early growth response protein 1) promoter region that is operatively linked to a gene encoding a fluorescent protein, or a gene encoding an siRNA directed against a constitutively expressed fluorescent protein, and controls expression of said gene; determining a level of fluid shear stress acting on cells during said bioprocess and/or during operation of said biotechnological device or system using said cell in real-time at any given time, and determining said level of fluid shear stress as the level of fluid shear stress that is acting on any cells subjected to said bioprocess and/or to operation of said biotechnological device or system at said given time.

    CELL-BASED SHEAR STRESS SENSOR
    4.
    发明公开
    CELL-BASED SHEAR STRESS SENSOR 审中-公开

    公开(公告)号：EP4279592A2

    公开(公告)日：2023-11-22

    申请号：EP23171982.4

    申请日：2023-05-08

    申请人： Sartorius Stedim Biotech GmbH

    发明人： Akbari, Samin ,  Leroux, Ann-Cathrin ,  Kwon, Taehong ,  Zehe, Christoph ,  Pollard, David

    IPC分类号： C12N15/113 ,  C12N15/65

    摘要： The present invention relates to isolated nucleic acid molecules, comprising a functional mammalian EGR-1 (early growth response protein 1) promoter region that is operatively linked to a gene encoding either a fluorescent protein or a gene encoding an siRNA directed against a constitutively expressed fluorescent protein and controls expression of said gene. The present invention further relates to nucleic acid vectors, comprising said nucleic, cells comprising said nucleic acids or vectors, being capable of sensing and indicating fluid shear stress acting on themselves, and methods of generating the same. Furthermore, the present invention relates to methods of evaluating fluid shear stress acting on cells in real-time during the operation of a biotechnological device or system, methods of evaluating a biotechnological device or system with respect to fluid shear stress acting on cells caused by operation of said device or system, and methods of designing a biotechnological device or system, said methods using said cells.

    METHOD FOR IN VITRO REPROGRAMMING OF FIBROBLASTS INTO SERTOLI CELLS AND APPLICATION THEREOF
    8.
    发明公开
    METHOD FOR IN VITRO REPROGRAMMING OF FIBROBLASTS INTO SERTOLI CELLS AND APPLICATION THEREOF 审中-公开

    公开(公告)号：EP3805375A1

    公开(公告)日：2021-04-14

    申请号：EP18925596.1

    申请日：2018-07-03

    申请人： Tsinghua University ,  Everlife Biomed Co. Ltd.

    发明人： KEE, Kehkooi ,  LIANG, Jianlin

    IPC分类号： C12N5/10 ,  C12N15/867 ,  C12N5/071 ,  C12N15/65

    摘要： Disclosed is a method for in vitro reprogramming of fibroblasts into Sertoli cells and an application thereof. The method includes the following steps: introducing a fluorescent protein reporter system specifically expressed in the Sertoli cells into the fibroblasts; followed by introducing a substance that increases the expression level and/or activity of the NR5A1 protein; adding G418 and culturing for 3-7 d; discarding the liquid phase, adding a medium for culturing fibroblasts and continuing culturing for 3-7 d; and isolating the Sertoli cells from the culture system. Experiments showed that cells prepared by the method fully acquire the characteristics of Sertoli cells, such as attracting endothelial cells, forming lipid droplets, interacting with germ cells, inhibiting lymphocyte growth and interleukin production and having immune privilege. The disclosure has application value in the fields of infertility treatment, allogeneic transplantation, and cell therapy.