EXPANSION OF MEGAKARYOCYTES AND IN VITRO THROMBOPOIESIS.

巨核细胞的扩增和体外血小板生成。

• 批准号: 7922604
• 负责人: Diana Gilligan
• 金额: $ 42.21万
• 依托单位: BLOODWORKS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7922604
• 关键词: Allogenic; Alloimmunization; Antibodies; Apoptosis; Autologous; Biological Assay; Blood Platelets; Blood donor; Bone Marrow; Cell Line; Cell surface; Characteristics; Chemicals; Clinical; Coculture Techniques; Collagen; Complex Mixtures; Consumption; Culture Media; Cytoskeleton; Data; Doctor of Medicine; Ear; Electron Microscopy; Endothelial Cells; Flow Cytometry; Genes; Goals; Hematopoietic; Hemorrhage; Hemostatic Agents; Human; Immunologics; In Vitro; Infusion procedures; Intensive Care; Intrinsic factor; Isotope Labeling; Lead; Life; Ligands; Light; Mass Spectrum Analysis; Measures; Medicine; Megakaryocyte Proliferation; Megakaryocytes; Methods; Microscopic; Nuclear; Operative Surgical Procedures; Oryctolagus cuniculus; Patients; Phenotype; Platelet Function Tests; Ploidies; Production; Proteins; Relative (related person); Research; Research Personnel; Risk; Ristocetin; SU 6656; Safety; Shapes; Signal Transduction Inhibitor; Signal Transduction Pathway; Staging; Staining method; Stains; Stem cell transplant; Supportive care; Surgical incisions; Techniques; Testing; Thrombin; Thrombocytopenia; Thrombopoiesis; Transfusion; Transplantation; Trauma; Umbilical Cord Blood; Viral; Whole Blood; annexin A5; base; cancer therapy; chemotherapy; cytokine; deprivation; gene repression; immortalized cell; immunogenicity; improved; in vivo; inhibitor/antagonist; irradiation; kinase inhibitor; knock-down; nonhuman primate; notch protein; particle; pathogen; progenitor; programs; response; src-Family Kinases; volunteer

The overall goal of this project is to develop methods for the production of functional platelets in vitro that could be used for clinical transfusion. Transfusion of allogeneic platelets is a critical part of supportive therapy for patients undergoing chemotherapy, stem cell transplantation, and those with thrombocytopenia due to increased consumption or inadequate production. However, the supply of transfusable platelets is dependent on volunteer donors. Furthermore, platelets have a short shelf-life and may be contaminated with bacterial or viral pathogens. Our proposal is based on the following hypotheses: 1) rational modulation of extrinsic and intrinsic factors will allow expansion of primary megakaryocytes in a manner that maintains an immature phenotype; 2) changes in culture conditions will facilitate terminal maturation and platelet shedding of the expanded MKs; and 3) platelet-like fragments, formed under these conditions, will have hemostatic activity, comparable to normal platelets. Specific Aims are: Aim 1: Develop methods for the expansion of megakaryocytes (MKs) from bone marrow or umbilical cord blood derived progenitors. Aim 2: Promote thrombopoiesis (i.e. fragmentation) of MKs and megakaryocyticcell lines in vitro. Aim 3: Test function of platelets produced in vitro. Successful completion of these Aims will lead directly to studies of in vitro derived platelets in non- human primates, with extensive testing to determine safety, efficacy, and platelet survival in vivo as well as immunogenicity upon repeated administration. This may eventually result in the production of autologous and/or donor-derived platelets from expanded progenitors in order to diminish alloimmunization, improve survival, and eliminate the risk of infectious complications.

该项目的总体目标是开发体外功能血小板生产的方法 可以用于临床输血。同种异体血小板的输血是 接受化疗，干细胞移植的患者的支持治疗 由于消费量增加或产生不足而导致的血小板减少症。但是，供应 可容纳的血小板取决于志愿捐助者。此外，血小板的保质期短 并可能被细菌或病毒病原体污染。我们的建议基于以下 假设：1）外在和内在因素的合理调制将允许主要的主要调制 巨核细胞以保持不成熟表型的方式； 2）培养条件的变化 将促进扩展的MK的终末成熟和血小板脱落； 3）类似血小板 在这些条件下形成的片段将具有与正常血小板相当的止血活性。 具体目的是： 目标1：开发从骨髓或 脐带血衍生的祖细胞。 目标2：在体外促进MK和巨核细胞系的血小板（即碎片化）。 AIM 3：体外产生的血小板的测试功能。 这些目标的成功完成将直接导致对非 - 非体外衍生血小板的研究 人类灵长类动物，并进行了广泛的测试，以确定体内的安全性，功效和血小板存活 作为重复给药时的免疫原性。这最终可能导致生产 自体和/或供体衍生的血小板从扩展的祖细胞中，以减少 同种免疫，改善生存和消除感染并发症的风险。