MEGAKARYOCYTE REAGENTS SPECIFIC FOR THEIR MATURATION

这项建议的总体目标是开发一种临床级巨核细胞发育介质(MDM)，支持人类巨核细胞的体外发育。使用这种独特的介质将提供有关这些难以捉摸但至关重要的细胞的几个关键间隙区域的信息；脂代谢、成熟调节和临床应用。创新的方面是增强脂质代谢的研究，细胞因子/基质分子组合的使用，以及体内巨核细胞移植模型的使用。MDM将在开发治疗血小板减少症和其他血小板疾病方面有临床应用，因为它允许产生大量的巨核细胞(&gt；10/9)，这是常规骨髓抽吸无法实现的。MDM还允许实验室间巨核细胞培养条件的标准化。在我们第一阶段研究的成功结果的基础上，我们在第二阶段提案中设计了相关的和高度关键的实验，以调查以下目标。具体目标1涉及MDM的进一步优化，特别侧重于增强巨核细胞的脂质代谢。特殊目的2研究作为复杂调控信号的特定细胞因子和基质分子的作用。具体目标3是测试和/或长期植入。这一目的也为基于细胞的血小板疾病治疗提供了一种临床前动物模型。具体目标4是将这些培养扩大到每个培养产生1-5x10/9巨核细胞的水平。这一第二阶段计划的成功完成将带来广泛使用的商业产品、临床相关的动物模型，以及进一步研究这些具有生物重要性的细胞的迫切需要的信息。 建议的商业应用： 支持巨核细胞增殖和分化的临床级试剂的开发将在开发治疗血小板减少症等造血疾病的新方法方面具有价值。

The overall objective of this proposal is to develop a clinical-grade Megakaryocyte Development Medium (MDM) that supports the ex vivo development of human megakaryocytes. Use of this unique medium will provide information on several critical gap-areas concerning these elusive yet critically important cells; lipid metabolism, regulation of maturation, and clinical utility. The innovative aspects are the studies on enhancing lipid metabolism, the use of cytokine/matrix molecule combinations, and an in vivo megakaryocyte-transplantation model. MDM will have clinical use in developing therapies for thrombocytopenia and other platelet disorders, in that it allows generation of large numbers of megakaryocytes (> 10/9) that are not possible via routine bone marrow aspiration. MDM also allows for the standardization of megakaryocyte culture conditions between laboratories. Building on the successful results of our Phase I studies, we have designed relevant and highly critical experiments in our Phase II proposal to investigate the following goals. Specific Aim 1 addresses the further optimization of MDM, with a particular focus on enhancing megakaryocyte lipid metabolism. Specific Aim 2 examines the effects of specific cytokines and matrix molecules as complex regulatory signals. Specific Aim 3 is to test and/or long-term engraftment. This aim also provides a pre-clinical animal model for cellular-based therapies of platelet disorders. Specific Aim 4 is to scale these cultures up to levels producing 1-5 x 10/9 megakaryocytes per culture. Successful completion of this Phase II proposal will result in a widely used commercial product, a clinically relevant animal model, and much needed information to further our studies on the development of these biologically important cells. PROPOSED COMMERCIAL APPLICATION: The development of clinical grade reagents that will support megakaryocyte proliferation and differentiation will be valuable in developing novel approaches for the treatment of hematopoietic disorders such as thrombocytopenia.