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    Creating novel hematopoietic cell lines by expressing altered retinoic acid receptors
    2.
    发明授权
    Creating novel hematopoietic cell lines by expressing altered retinoic acid receptors 失效
    通过表达改变的视黄酸受体来创建新的造血细胞系

    公开(公告)号：US5830760A

    公开(公告)日：1998-11-03

    申请号：US592383

    申请日：1996-01-26

    申请人： Schickwann Tsai ,  Steven J. Collins

    发明人： Schickwann Tsai ,  Steven J. Collins

    IPC分类号： C07K14/705 ,  C12N5/0789 ,  C12N5/16

    CPC分类号： C07K14/70567 ,  C12N5/0647 ,  C12N2501/125 ,  C12N2501/22 ,  C12N2501/23 ,  C12N2501/385 ,  C12N2510/00

    摘要： Methods for establishing continuous SCF dependent lympho-hematopoietic progenitor cell lines capable of differentiating into erythroid, myeloid, and B lymphocytic lineages, and GM-CSF dependent neutrophil progenitor cell lines capable of differentiating into neutrophils but not into monocytes, mast cells, or basophils, by introducing into bone marrow, fetal spleen, fetal liver, or other hematopoietic myeloid cells nucleic acid encoding a dominant negative suppressor of a retinoic acid receptor-alpha and a selectable marker, and culturing the recombinant cells in culture medium containing SCF or GM-CSF, agents allowing for selective growth of the recombinant cells, and a level of retinoic acid of less than about 10.sup.-8 M to about 10.sup.-9 M in the case of establishing neutrophilic progenitor cell lines. Addition of a retinol compound induces the latter cell line to differentiate into neutrophils.

    摘要翻译： PCT No.PCT / US94 / 08450 Sec。 371日期1996年1月26日 102（e）日期1996年1月26日PCT提交1994年7月28日PCT公布。 公开号WO95 / 04143 日期1995年2月9日建立能够分化为红细胞，骨髓和B淋巴细胞谱系的连续SCF依赖性淋巴造血祖细胞系和能够分化为嗜中性粒细胞而不分化为单核细胞的GM-CSF依赖性嗜中性粒细胞祖细胞系的方法 细胞或嗜碱性粒细胞，通过引入编码视黄酸受体-α和选择性标记的显性负抑制子的骨髓，胎儿脾脏，胎肝或其他造血骨髓细胞核酸，并在含有 在建立嗜中性粒细胞祖细胞系的情况下，SCF或GM-CSF，允许重组细胞的选择性生长的试剂和小于约10-8M至约10-9M的视黄酸水平。 加入视黄醇化合物诱导后一种细胞系分化为嗜中性粒细胞。

    Penumbra Nucleic Acid Molecules, Proteins and Uses Thereof
    3.
    发明申请
    Penumbra Nucleic Acid Molecules, Proteins and Uses Thereof 审中-公开
    半胱氨酸核酸分子，蛋白质及其用途

    公开(公告)号：US20090264354A1

    公开(公告)日：2009-10-22

    申请号：US11992872

    申请日：2006-09-28

    申请人： Schickwann Tsai ,  Zhong Chen

    发明人： Schickwann Tsai ,  Zhong Chen

    IPC分类号： A61K38/16 ,  C07K14/705 ,  C12Q1/68

    CPC分类号： C07K14/70596

    摘要： The present invention relates to murine and human Penumbra (for proerythroblast nu[new] Membrane) nucleic acid molecules, proteins and the uses thereof. The invention further relates to the use of Penumbra molecules for the detection of 7q31q32-related deletions, including such deletions associated with myeloid malignancies, particularly detection by hybridization using Penumbra-based probes.

    摘要翻译： 本发明涉及鼠和人类Penumbra（对于红细胞新生物）核酸分子，蛋白质及其用途。 本发明还涉及半影分子用于检测7q31q32相关缺失的用途，包括与骨髓恶性肿瘤相关的这种缺失，特别是使用基于半胱氨​​酸的探针进行杂交的检测。

    Low-macrophage-adhesion/activation culture devices for continuous hematopoiesis and expansion of hematopoietic stem cells and progenitor cells
    4.
    发明公开
    Low-macrophage-adhesion/activation culture devices for continuous hematopoiesis and expansion of hematopoietic stem cells and progenitor cells 审中-公开

    公开(公告)号：US20230159873A1

    公开(公告)日：2023-05-25

    申请号：US17971623

    申请日：2022-10-23

    申请人： Schickwann Tsai

    发明人： Schickwann Tsai

    IPC分类号： C12M1/00

    CPC分类号： C12M23/20 ,  C12M23/34

    摘要： Hematopoietic stem cells are extremely difficult to maintain or expand in vitro. Two observations in traditional long-term bone marrow cultures strongly suggest that macrophages may be at the root of the problem: First, micromolar concentrations of hydrocortisone improve the longevity of long-term bone marrow cultures and hydrocortisone is known as a potent inhibitor of macrophage production of pro-inflammatory cytokines, chemokines, enzymes, nitrogen oxide and reactive oxygen species and redirects macrophages to the anti-inflammatory differentiation pathway; Second, the decline of hematopoiesis in long-term bone marrow cultures coincides with the development of large numbers of adherent and non-adherent macrophages including foreign body giant cells. These adherent macrophages and foreign body giant cells exhibit well-spread morphology, contain numerous lysosomes and phagolysosomes in the cytoplasm and are metabolically active. We hypothesize that hydrocortisone fails to suppress all aspects of macrophage pro-inflammatory activation/differentiation, resulting in the production of inhibitors or toxins of hematopoiesis. Macrophage adhesion in cell culture depends on serum proteins pre-adsorbed to the tissue-culture-treated polystyrene (TC-PS), which adsorbs proteins via mostly hydrophilic interactions. TC-PS is used in almost all tissue culture devices currently available. Cellular adhesion provides a strong stimulus for metabolic, mitotic and certain gene activities. Therefore, we seek to reduce macrophage adhesion and activation by culturing bone marrow cells in tissue culture devices composed of or covered with polymers with very different protein-binding characteristics than TC-PS such as polyethylene (PE) and other polyolefins, the latter bind proteins via exclusively hydrophobic interactions. As a result, polyolefins bind different proteins and in lower quantities than TC-PS. Furthermore, PE does not contain additional chemical features like the phenolic rings of polystyrene that might contribute to protein binding and macrophage adhesion/activation. Using these new culture devices, we developed a drastically different long-term bone marrow culture, the “Low Macrophage-Adhesion/Activation” (LoMAC) bone marrow culture. In LoMAC bone marrow culture, hematopoiesis continues for months to over a year and hematopoietic stem cells are amplified gradually. In stark contrast to traditional long-term bone marrow cultures, de novo erythropoiesis and megakaryocytopoiesis proceed robustly in the LoMAC bone marrow culture and B-lymphocyte and natural killer cell progenitors can be continuously derived. Thus, these new culture devices and the associated LoMAC culture method offer a new way to study hematopoiesis in vitro and provide a more robust platform for the expansion of hematopoietic stem cells and progenitors ex vivo.

    Anti-penumbra monoclonal antibodies for detection and therapy of normal and abnormal B lymphocytes
    5.
    发明申请
    Anti-penumbra monoclonal antibodies for detection and therapy of normal and abnormal B lymphocytes 审中-公开

    公开(公告)号：US20180362660A1

    公开(公告)日：2018-12-20

    申请号：US15436855

    申请日：2017-02-19

    申请人： Schickwann Tsai

    发明人： Schickwann Tsai

    IPC分类号： C07K16/36 ,  C07K16/30

    摘要： Penumbra is the newest member of the tetraspanin superfamily of membrane proteins. A major obstacle in penumbra research has been the lack of monoclonal antibodies against the native penumbra. In this invention, we detail the establishment and characterization of monoclonal antibodies that recognize both human and mouse penumbras on living cells. Furthermore, we created chimeric mouse-human IgG1 antibodies from these mouse monoclonal antibodies. Using these antibodies, we demonstrate for the first time that penumbra is expressed on the surface of virtually all CD19+ or CD20+ B lymphocytes in blood, bone marrow, spleen and lymph nodes. In vivo, these monoclonal antibodies shrank lymphoid follicles in spleen. Thus, these antibodies establish penumbra as a novel B cell marker with a wider range of expression level than CD19 or CD20. These monoclonal antibodies pave the way for new research and potential therapeutic applications in immunology, hematology and oncology.