In vitro bioreactor sys for platelet formation

描述(申请人提供)：每年进行数百万次的血小板输注，但这种血液成分的供应有限，因此患者治疗疾病的机会是有问题的。也有许多疾病的血小板生成或功能受损，导致严重后果，临床选择有限。为了解决目前的这些局限性，体外产生功能性血小板的新模式将为许多患者提供重大好处，并提供一种方法，使系统研究参与功能性血小板形成的机制成为可能。我们在这个项目中的目标是在我们最近成功的研究的基础上，设计出一种新的生物反应器系统来容纳巨核细胞(MKs)并在体外产生功能性血小板。我们将利用这一系统来解决我们的假设；体外工程微环境可以被定制来优化功能性血小板的形成。为了满足计划中的需求，我们将：(1)建立一个基于生物反应器的3D组织系统，以研究MK的发育和血小板释放的机制，并根据系统产生和恢复的功能性血小板来研究功能结果；(2)使用该3D组织系统，通过包括来自血小板相关疾病患者的MK，来研究与生化信号和环境成分(即基质、氧分压)相关的血小板产生以及病理性巨核生成的机制。有了这个系统的洞察，我们将能够询问来自正常来源和疾病来源的MK的成熟度，以便开始确定MK结果(黏附、迁移、原血小板形成、血小板产生和功能)的差异。这项拟议研究的结果将是双重的：(1)一个新的实验室模型，用于研究正常和异常(疾病)状态下MK的发育、血小板原形成和血小板释放，以及(2)对这些过程的机械洞察。从长远来看，建立这一新的体外组织系统将允许更系统地了解MK发育过程，以及洞察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.