Bioreactor Core Facility

生物反应器核心设施 导言和具体目标 组织工程（TE），即，活的、三维的、功能性组织类似物的制造是一种快速的、 开发有可能彻底改变医疗保健的研究领域。工程化三维组织 类似物旨在替代组织;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间相互作用。 质量传输限制的重要性随着TE构造的尺寸的增加而迅速增长;在静态情况下， 培养时，它可能在超过几mm 3的构建体尺寸时变得受限。在这一点上，生物反应器变得必不可少 提供一个环境，在其中营养供应和废物去除，而且环境条件， 例如pH、温度或压力，可以被监测和控制。 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 并且操作复杂，并且对于给定TE应用的最佳设计构成几乎没有共识。 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，他们正在设计或购买自己的设备。此外，TE现在正处于测试阶段， 一系列实验条件的变化变得至关重要。及时完成此类实验 将需要大量的反应堆。这为建立生物反应器提供了明确的依据 核心贷款，即，实验室致力于提供最先进的技术，仪器和专业的 发展支持，以及对教职员工和学生的专门培训。 本计划项目补助金中提出的所有项目都将在某个时候使用生物反应器。 凯斯西储大学目前没有正式的生物反应器核心设施。这个PI 核心设施和他的同事已经开发了一个模块化的生物反应器系统，以解决软骨的需要， 组织工程学核心反应堆设计简单，但可高度扩展，以允许广泛的治疗 和监测方式。除了本项目建议书中详细介绍的项目外， 在其他资助项目中得到了证明。这在技术、科学和经济上都是有意义的 集中这些项目的所有生物反应器相关步骤，从实验设计到设计和建造 生物反应器的硬件，到实际的生物反应器操作。此外，这将使调查人员能够集中精力 更多地关注他们项目的核心基础科学问题。 拟议的核心设施将强调服务和核心设施的研究组成部分， 具体目标如下： 具体目标I：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： <$最先进的生物反应器培养设施，供计划项目研究人员进行常规组织工程 技术专长-这一核心设施将为所有生物反应器相关领域提供训练有素的技术人员。 因此，它提供了最好的技术专长。协助实验设计， 基于对系统的能力和局限性的了解，以及特定项目的开发， 根据需要扩展现有系统。 具体目标II：开发一个临床上有用的骨软骨组织工程工具集。本研究 组件将专注于开发系统的要素，以确保临床可扩展性并最大限度地提高临床 效用这些包括过程监控、数据采集和控制。具体目标 包括以下步骤： 通过在线监测环境参数和反馈控制实现生物反应器介质的稳态 通过中等流速和/或通过使用缓冲液。 开发MRI方案，以非侵入性方式监测植入物发育。 <$对整个生物反应器系统进行详细的计算机建模，包括流体力学 以及与构建体的气体/营养交换。 开发一次性版本的介质润湿生物反应器组件，用于未来的临床应用。 核心设施将提供所有这些服务和专业知识，使Pis项目能够专注于实验性研究。 设计和结果解释。核心将由细胞，生物力学和形态学补充 核心，并将受益于收集有关需求的关键信息 通过完善不同实验类型的计算机模型模板，这些数据 然后将从单一来源提供，以帮助核心用户设计他们的实验。因此，我们预计 这一设施将成为许多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.