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    • 1. 发明申请
    • IN VITRO GROWTH OF NEUTROPHIL AND MEGAKARYOCYTE PRECURSORS IN SERUM-FREE MEDIA
    • 在无血清培养基中的中性粒细胞和巨细胞前体的体外生长
    • WO1995006112A1
    • 1995-03-02
    • PCT/US1994009622
    • 1994-08-23
    • BAXTER INTERNATIONAL INC.
    • BAXTER INTERNATIONAL INC.SMITH, Stephen, L.QIAO, XiaoyingMACIUKAS, Susan, M.BENDER, James, G.VAN EPPS, Dennis, E.LOUDOVARIS, Maureen, F.
    • C12N05/08
    • C12N5/0644A61K35/12C12N5/0642C12N2500/25C12N2500/36C12N2500/44C12N2500/90C12N2500/98C12N2501/125C12N2501/22C12N2501/23C12N2501/39
    • The invention provides serum-free, animal protein-free media formulations to be used in conjunction with hematopoietic growth factors for the in vitro growth of human neutrophil and megakaryocyte precursors. The medium is comprised of a base medium, corticosteroid, transferrin, insulin, cholesterol, ethanolamine, and human albumin. The invention also provides methods for preparing serum-free, animal protein-free suspensions of human hematopoietic precursor cells wherein the cellular component comprises at least about 16 % neutrophil precursors and at least about 1 % megakaryocyte precursors. Serum-free, animal protein-free suspensions of human hematopoietic cells are provided wherein the cellular component comprises at least about 30 %, preferably greater than 60 % neutrophil precursors. The neutrophil precursors are comprised of blast cells, promyelocytes, neutrophilic myelocytes, and neutrophilic metamyelocytes. Also provided are serum-free, animal protein-free cell suspensions wherein the cellular component comprises at least about 3 %, preferably greater than 8 % megakaryocyte precursors. Also provided are serum-free, animal-protein free cell suspensions wherein the cellular component comprises colony-forming units and cluster-forming units.
    • 本发明提供了与造血生长因子结合使用的无血清,无动物蛋白质的培养基制剂,用于人嗜中性粒细胞和巨核细胞前体的体外生长。 培养基由基础培养基,皮质类固醇,转铁蛋白,胰岛素,胆固醇,乙醇胺和人白蛋白组成。 本发明还提供了制备人类造血前体细胞的无血清,无动物蛋白质的悬浮液的方法,其中细胞组分包含至少约16%的嗜中性粒细胞前体和至少约1%的巨核细胞前体。 提供人造血细胞的无血清,不含动物蛋白的悬浮液,其中细胞组分包含至少约30%,优选大于60%的嗜中性粒细胞前体。 中性粒细胞前体由胚细胞,前髓细胞,嗜中性粒细胞和嗜中性粒细胞组成。 还提供了无血清,无动物蛋白的细胞悬浮液,其中细胞组分包含至少约3%,优选大于8%的巨核细胞前体。 还提供了无血清,无动物蛋白质的细胞悬浮液,其中细胞组分包含集落形成单位和簇形成单位。
    • 4. 发明申请
    • DENDRITIC CELLS PRODUCED IN SERUM-FREE CULTURE
    • 在无血清培养物中生产的细胞
    • WO1998006826A1
    • 1998-02-19
    • PCT/US1997014383
    • 1997-08-13
    • BAXTER INTERNATIONAL INC.
    • BAXTER INTERNATIONAL INC.VACHULA, MonaALZONA, Mortimer, T.AONO, Frederick, M.KOWALKOWSKI, Kenneth, L.SMITH, Stephen, L.UNVERZAGT, Kristen, L.VAN EPPS, Dennis, E.
    • C12N05/08
    • C12N5/0639A61K38/00A61K2039/5154A61K2039/5158C12N2500/05C12N2500/14C12N2500/36C12N2500/38C12N2500/90C12N2501/01C12N2501/02C12N2501/125C12N2501/22C12N2501/25C12N2501/515
    • A method for producing human dendritic cells (DC) for therapeutic purposes using serum-free, animal protein-free medium. The method involves culturing mononuclear cells from peripheral or umbilical cord blood or bone marrow of an individual human donor in a culture container having at least one inner surface of polystyrene in the above described medium containing at least one hematopoietic growth factor, and maintaining the culture to produce a culture enriched for DC. The DC enrichment is evidenced by at least about 10% of the total cells in the culture exhibiting DC processes, or at least 2.5 % of the total cells expressing at least one DC antigen such as CD80, CD86, or CD1a. The culture method is preferably carried out in a gas-permeable container in a closed fluid path system. The DC so produced can be pulsed with an antigen such as a viral, bacterial, tumor, or cell surface antigen, and the resulting primed dendritic cells can be co-cultured with T cells to produce antigen-specific human T-cells for therapeutic use. Useful for treatment of viral or bacterial infections, useful as a cancer vaccine, useful to induce tolerance when transplant of allo- or xeno-graft cells or organ is intended.
    • 用于使用无血清,无动物蛋白质的培养基用于治疗目的的人树突状细胞(DC)的制备方法。 该方法包括在含有至少一种造血生长因子的上述培养基中具有聚苯乙烯的至少一个内表面的培养容器中培养单个人供体的外周或脐带血或骨髓的单核细胞,并将培养物保持 生产富含DC的培养基。 DC富集由显示DC过程的培养物中的总细胞的至少约10%或表达至少一种DC抗原如CD80,CD86或CD1a的总细胞的至少2.5%来证明。 培养方法优选在封闭流体路径系统中的透气性容器中进行。 如此产生的DC可用诸如病毒,细菌,肿瘤或细胞表面抗原的抗原脉冲,并且所得引发的树突状细胞可与T细胞共培养以产生用于治疗用途的抗原特异性人T细胞 。 用于治疗病毒或细菌感染,可用作癌症疫苗,用于诱导同种异体移植或异种移植细胞或器官移植时的耐受性。