Bioreactor Core Facility

生物反应器核心设施 引言和具体目标 组织工程(TE)，即制造活的、三维的、有功能的组织类似物，是一种快速的 开发有可能给医疗保健带来革命性变化的研究领域。工程化三维组织 类似物旨在取代组织；此外，它们还可以作为研究组织的研究工具 形态发生和复杂的细胞-细胞相互作用。 随着TE结构尺寸的增加，质量传输限制的重要性迅速增加；在静态情况下 培养，它可以成为限制在结构大小超过几个毫米米。在这一点上，生物反应器变得至关重要。 提供一个提供养分供应和废物清除的环境，但也提供环境条件，如 因为pH、温度或压力是可以监测和控制的。 几种生物反应器设计已经商业化；然而，这些装置仍然相当昂贵。 而且操作复杂，对于什么构成给定TE应用的最佳设计，几乎没有共识。 希望开发大规模TE结构作为其研究计划一部分的研究人员必须 目前设计或购买自己的设备。此外，TE现在正在达到一个阶段，即测试 实验条件的阵列正在变得至关重要。及时完成这样的实验 将需要大量的反应堆。这为建立生物反应器提供了明确的理由 核心设施，即致力于提供最先进的技术、仪器和专业技术的实验室 发展支持，并对教职员工和学生进行专门培训。 本计划中提出的所有项目都将在某一时刻使用生物反应器。 凯斯西储大学目前没有正式的生物反应器核心设施。这件事的PI 核心设施和他的同事开发了一种模块化生物反应器系统，以满足软骨的需求 组织工程学。堆芯反应堆设计简单，但可高度扩展，以允许广泛的处理 和监控模式。除本计划项目建议书中详述的项目外，本系统 在其他资助项目的背景下证明了自己。它在技术上、科学上和经济上都有意义 集中这些项目与生物反应器有关的所有步骤，从实验设计到设计和施工 生物反应器硬件，到实际的生物反应器操作。此外，这将使调查人员能够专注于 他们将更多的注意力放在对他们的项目至关重要的基础科学问题上。 拟议的核心设施将强调总结的服务和核心设施研究组成部分 在以下具体目标中： 具体目标一：为每个PPG项目提供全方位服务的研究生物反应器设施，包括： ？为常规组织工程项目研究人员提供最先进的生物反应器培养设施 ？技术专长-这一核心设施将为所有与生物反应器有关的人员提供训练有素的熟练人员 业务，从而提供尽可能好的技术专长。实验设计方面的协助， 基于对系统能力和局限性的了解，以及特定于项目的开发， 将为现有系统提供需求驱动的扩展。 具体目的二：开发一套临床实用的骨软骨组织工程工具箱。这项研究 组件将专注于开发系统的元素，以确保临床可伸缩性并最大限度地提高临床 实用程序。其中包括过程监控、数据采集和控制。应针对的具体方面 包括： ？生物反应器介质动态平衡，通过在线监测环境参数和反馈控制 通过介质流速和/或通过使用缓冲器。 开发磁共振成像方案，以非侵入性方式监测种植体的发育。 ？对整个生物反应器系统进行详细的计算机模拟，包括水动力学 以及与建筑的气体/养分交换。 ？为未来的临床应用开发一次性使用的湿式生物反应器组件。 核心设施将提供所有这些服务和专业知识，使PIS项目得以专注于试验性 设计和结果解释。核心将由细胞、生物力学和形态学补充 本提案中其他地方概述的核心，并将从收集有关需求的关键信息中受益 通过完善不同实验类型的计算机模型模板，研究了组织类型。这些数据 然后将从单一来源获得，以帮助核心用户设计他们的实验。因此，我们预计 这一设施将成为大量调查人员的宝贵资源。

Bioreactor Core Facility Introduction and Specific Aims Tissue engineering (TE), i.e., the fabrication of living, 3-dimensional, functional tissue analogs, is a rapidly developing research area that has the potential to revolutionize healthcare. Engineered three-dimensional tissue analogs are intended to replace tissue; in addition, they can serve as research tools to investigate tissue morphogenesis and complex cell-cell interactions. The significance of mass-transport limitations grows rapidly with increasing size of a TE construct; in static culture it can become limiting at construct sizes over a few mm3. At this point, a bioreactor becomes essential to provide an environment in which nutrient supply and waste removal, but also environmental conditions such as pH, temperature, or pressure, can be monitored and controlled. Several bioreactor designs have been commercialized; however, these devices remain quite expensive and complex to operate, and there is little consensus as to what constitutes the best design for a given TE application. Investigators wishing to develop large-scale TE constructs as part of their research program must currently design or purchase their own equipment. Furthermore, TE is now reaching a stage in which the testing of arrays of experimental conditions is becoming critical. Completing such experiments in a timely fashion will require a large number of reactors. This provides a clear rationale for the establishment of a Bioreactor Core Facility, i.e., a laboratory dedicated to providing state-of-the-art technical, instrument, and professional development support, and specialized training of faculty, staff and students. All the projects proposed in this Program Project Grant will make use of a bioreactor at one point or another. Case Western Reserve University does not currently have a formal bioreactor core facility. The PI of this core facility and his co-workers has developed a modular bioreactor system to address the needs of cartilage tissue engineering. The core reactor design is simple, but highly extensible, to allow for a wide range of treatment and monitoring modalities. In addition to the projects detailed in this Program Project proposal, this system has proven itself in the context of other funded projects. It makes technical, scientific and economic sense to centralize all bioreactor-related steps of these projects, from experimental design, to design and construction of bioreactor hardware, to the actual bioreactor operation. Further, this will allow the investigators to focus more of their attention on the basic science questions that are central to their projects. The proposed core facility will emphasize service and core facility research components that are summarized in the following Specific Aims: Specific Aim I: to provide a full-service research bioreactor facility to each PPG project, including: ¿ State-of-the-art bioreactor culture facilities for Program Project investigators for routine tissue engineering ¿ Technical expertise - this core facility would provide trained and skilled personnel for all bioreactor related operations, thus providing the best possible technical expertise. Assistance in experimental design, based on knowledge of the capabilities and limitations of the system, and development of projectspecific, need-driven extensions to the existing system, will be provided. Specific Aim II: to develop a clinically useful toolset for osteochondral tissue engineering. This research component will focus on developing elements of a system that will ensure clinical scalability and maximize clinical utility. These include process monitoring, and data acquisition and control. Specific aspects to be targeted include: ¿ Bioreactor medium homeostasis, by online monitoring of environmental parameters and feedback control through medium flow rate and/or through the use of buffers. ¿ MRI protocol development to monitor implant development non-invasively. ¿ Implementation of detailed computer modeling of the bioreactor system as a whole, including hydrodynamics and gas/nutrient exchange with the construct. ¿ Development of disposable versions of the media-wetted bioreactor components for future clinical applications. The core facility will provide all these services and expertise, freeing the Project Pis to focus on experimental design and result interpretation. The core will be complemented by the Cell, Biomechanics, and Morphology cores outlined elsewhere in this proposal, and will benefit by gathering critical information on the requirements of tissue type investigated, by perfecting computer model templates for different experiment types. These data will then be available from a single source to aid core users in designing their experiments. We therefore anticipate that this facility will become an invaluable resource to a significant number of investigators.