ShEEP application for Integrated Hypoxia Exposure and Analysis Core

这项提议的主要目标是建立一个综合缺氧细胞核心设施(HCCF)，作为 拉尔夫·H·拉尔夫研究所现有的细胞和分子评估核心(CMEC)设施。 约翰逊退伍军人医疗中心(RHJVAMC)。气候变化框架将以三个左右为中心。 最先进的低氧低氧工作站，每个都具有新的功能，包括 集低氧细胞培养孵化器和处理站于一身。这将允许长期 细胞在单一可控氧、二氧化碳和氮气环境中的传代和处理。 此外，作为全面服务核心，HCCF将包括一个受控速率的细胞冷冻系统，以 规范细胞冷冻，降低变异性，优化加工、冷冻、储藏、解冻 回收有价值的小鼠和人类细胞。这一核心设施的总体目标是提供 必要的工具和服务，不仅是对需要低氧细胞培养的领域进行尖端研究的关键 对于RHJVAMC的干细胞、癌症和缺血性损伤研究项目，但这是针对 正在出现的问题，被认为是所有体外和原位研究的障碍，即生理上的 用于细胞和组织加工、培养和研究的氧气环境。现在可以理解的是 标准的常氧或室内空气组织培养技术是非生理学的，在以下方面会产生问题 对研究数据的解释。这也符合退伍军人管理局和美国国立卫生研究院最近发出的紧急呼吁，要求发展 适当的生理上相关的“正常”和疾病模型系统，以防止资源浪费在 未能模拟真实世界生理学的实验，使研究文献变得混乱。因此，可用性 低氧细胞核心设施的建立将不仅允许与细胞和组织的病理减少相关的研究 氧气见于疾病和损伤中，但它将增强退伍军人管理局调查人员执行“标准”组织的能力 以更具生理相关性的方式培养文化，并支持和推进直接影响 我们退伍军人的健康。我们的调查人员已经确定了四个主要目标，这些目标将通过 开创性技术和集成核心：1)需要低氧细胞培养仪器，以允许 长期培养通常存在于“低氧”微环境中的干细胞；2)需要 了解病理性低氧对组织/细胞损伤的影响(例如炎症和缺血性损伤，包括 中风和心肌梗死)，以及在肿瘤中发现的不同缺氧和缺氧区的建模；3) 允许将现有或未来的CMEC仪器放置在缺氧室中，以实时检测活细胞/组织 分析；以及4)改进了难以冷冻和恢复的细胞系的处理、储存和回收， 特别是人类干细胞和原代细胞。关键是，这一核心将填补RHJVAMC尚未满足的主要需求 因为在退伍军人管理局或其附属机构内没有可识别的低氧细胞培养系统， 南卡罗来纳医科大学。最先进的低氧细胞培养技术 HypOxygen工作站和其他核心元素以及在已建立的 Va CMEC将大大加强我们空间站的研究能力，促进基础科学发现 生理和病理过程的潜在机制，以及这些发现的翻译以支持 快速实施尖端个性化医疗，提高我国医疗质量 国家退伍军人，这是国务卿卓越蓝图和部长舒尔金的蓝图中概述的具体指令 10点计划。

The primary goal of this proposal is to establish an integrated Hypoxia Cell Core Facility (HCCF) as part of the existing Cellular and Molecular Evaluation Core (CMEC) Facility within Research Service at the Ralph H. Johnson VA Medical Center (RHJVAMC). The HCCF will be centered around three state-of-the-art HypOxygen hypoxia workstations, each with novel capabilities, that incorporate integrated hypoxia cell culture incubators and processing stations in one unit. This will permit long-term passaging and treatment of cells in a single controlled oxygen, CO2 and nitrogen environment. Additionally, as a full-service Core the HCCF will include a controlled rate cell freezing system to standardize cell freezing, reduce variability and optimize the processing, freezing, storage and thawing recovery of valuable murine and human cells. The overarching objective of this core facility is to provide tools and services necessary to not only for cutting edge research into areas requiring hypoxia cell culture critical for stem cell, cancer and ischemic injury research programs at the RHJVAMC, but that are addressing an emerging issue that is being recognized as a barrier for all in vitro and in situ research, namely physiologically oxygen environments for the processing, culturing and study of cells and tissues. It is now understood that standard “normoxic” or room air tissue culture techniques are non-physiologic and create problems in terms of interpretation of research data. This also meets the recent and urgent VA and NIH calls for development of appropriate physiologically relevant “normal” and disease model systems to prevent the wasting of resources on experiments that fail to model real world physiology and muddy the research literature. Therefore, the availability of a Hypoxia Cell Core Facility will not only permit studies related to pathological reductions of cell and tissue oxygen seen in disease and injury, but it will enhance the ability of VA investigators to perform “standard” tissue culture in a more physiologically relevant manner and support and advance research that directly impacts the health of our Veterans. Our investigators have identified four primary aims that will be accomplished through this groundbreaking technology and integrated Core: 1) The need for hypoxia cell culturing instrumentation to allow long-term culturing of stem cells which normally reside in “hypoxic” niches microenvironments.; 2) The need to access the effects of pathological low oxygen in tissue/cell injury (e.g. inflammation, and ischemic injury including stroke and myocardial infarction), and for modeling of the different hypoxic and anoxic zones found in tumors; 3) allowing existing or future CMEC instruments to be placed within a hypoxic chamber for live cell/tissue real-time analysis; and 4) Improved processing, storage and recovery of difficult to freeze and recover cell lines, in particular human stem cells and primary cells. Critically, this core will fill a major unmet need at the RHJVAMC in that there are no identifiable hypoxia cell culturing systems available either within the VA or its affiliate, the Medical University of South Carolina. The state-of-the-art hypoxia cell culturing technology afforded by HypOxygen workstations and other Core elements together with the addition of the HCCF into the established VA CMEC will greatly strengthen the research capabilities of our station, facilitating basic scientific discovery of mechanisms underlying physiological and pathological processes, and translation of these findings to support the rapid implementation of cutting-edge personalized medical treatment to improve quality healthcare for our Nation’s Veterans, a specific directive outlined in the Secretary’s Blueprint for Excellence and Secretary Shulkin’s 10-point plan.