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    Method of differentiating human pluripotent stem cells into primitive macrophages
    14.
    发明授权
    Method of differentiating human pluripotent stem cells into primitive macrophages 有权

    公开(公告)号：US10724003B2

    公开(公告)日：2020-07-28

    申请号：US15544254

    申请日：2016-01-18

    Applicant: AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH

    Inventor： Hwei-In Shawn Lim ,  Tara Huber ,  Florent Ginhoux

    IPC: C12N5/00 ,  C12N5/0786 ,  A61K35/15 ,  G01N33/50

    Abstract: The present invention relates to a method of culturing primitive-like macrophages from stem cells, a kit when used in the method thereof and uses of the primitive like macrophage for in-vitro disease models and for screening compounds for therapy. One embodied culture method comprises contacting and incubating embryonic stem cells or induced pluripotent stem cells with a serum-free culture media comprising a GSK3 inhibitor to differentiate stem cells into cells of the mesoderm lineage, followed by incubation with a culture media comprising Dickkopf-related protein 1 (DKK1) to differentiate the mesoderm into cells of hematopoietic lineage, maturing hematopoietic cells and incubating these cells with a culture media comprising M-CSF to drive differentiation into primitive-like macrophages. Another embodiment comprises incubating the stem cells with serum-free culture media comprising FGF2 and BMP4 to induce differentiation into cells of the mesoderm lineage, followed by incubating the cells with a culture media comprising FGF2, BMP4, Activin A and VEGF to differentiate the cells of the mesoderm lineage into cells of the hematopoietic cell lineage, maturing the cells of the hematopoietic cell lineage and lastly, incubating the matured hematopoietic cells with culture media comprising M-CSF to drive the differentiation of hematopoietic cells into primitive-like macrophages.

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