In vitro bioreactor sys for platelet formation

描述（由申请人提供）：每年进行数百万次血小板输注，但这种血液成分的供应有限，因此患者获得治疗疾病是有问题的。还有许多疾病，其中血小板产生或功能受损，导致严重后果，并且可用的临床选择有限。为了解决这些目前的局限性，在体外产生功能性血小板的新模式将为许多患者提供主要益处，并提供一种方法来允许系统研究功能性血小板形成中涉及的机制。我们在这个项目中的目标是建立在我们最近成功的研究中，一种新的生物反应器系统被设计成容纳巨核细胞（Mks），并在体外产生功能性血小板。我们将利用这个系统来解决我们的假设;体外工程微环境可以定制，以优化功能性血小板的形成。为了满足该计划的需求，我们将：（目的1）建立基于生物反应器的3D组织系统以研究Mk发育和血小板释放的机制，以及从系统产生和回收的功能性血小板的功能结果，以及（目的2）使用该3D组织系统研究与生化信号传导和环境组分相关的血小板产生机制（即基质，氧张力）以及病理性巨核细胞生成，通过包括来自患有血小板相关疾病的患者的Mk。通过对该系统的深入了解，我们将定位于询问来自正常与患病来源的Mk的成熟，以便开始确定Mk结局（粘附、迁移、前血小板形成、血小板生成和功能）的差异。拟议研究的结果将是双重的：（1）在正常和异常（疾病）状态下Mk发育、前血小板形成和血小板释放的新实验室模型，以及（2）对这些过程的机制性见解。从长远来看，建立在这种新的体外组织系统上将允许更系统地了解Mk发育所涉及的过程，以及深入了解干预与这些细胞相关的疾病状态的模式。

DESCRIPTION (provided by applicant): Millions of platelet transfusions are conducted each year, yet the supply of this blood component is limited, thus patient access to treat disorders is problematic. There are also many diseases where platelet production or function are impaired, resulting in severe consequences and where there are limited clinical options available. To address these current limitations, new modes to generate functional platelets in vitro would provide a major benefit to many patients, as well as provide an approach to permit the systematic investigation of mechanisms involved in functional platelet formation. Our goal in this Project is to build upon our recent successful studies where a novel bioreactor system was engineered to house megakaryocytes (Mks) and to generate functional platelets in vitro. We will exploit this system to address our hypothesis; engineered microenvironments in vitro can be tailored to optimize the formation of functional platelets. To address the needs in the program we will: (Aim 1) establish a bioreactor-based 3D tissue system to study the mechanisms of Mk development and platelet release, with functional outcomes in terms of functional platelets generated and recovered from the system, and (Aim 2) to use this 3D tissue system to investigate mechanisms of platelet production related to biochemical signalling and environmental components (i.e. matrices, oxygen tension) as well as pathological megakaryopoiesis by including Mks from patients with platelet related diseases. With insight from this system we will be positioned to interrogate the maturation of Mks from both normal vs. diseased sources in order to begin to establish differences in Mk outcomes (adhesion, migration, proplatelet formation, platelet production and function). The outcome for the proposed study would be twofold: (1) a new laboratory model for Mk development, proplatelet formation and platelet release in normal and abnormal (disease) states, and (2) mechanistic insight into these processes. In the long run, building upon this new in vitro tissue system would allow for the more systematic understanding of the processes involved in Mk development, as well as insight into modes to intervene in disease states associated with these cells.