ShEEP application for Integrated Hypoxia Exposure and Analysis Core

用于集成缺氧暴露和分析核心的 ShEEP 应用

• 批准号: 9795680
• 负责人: Donald R. Menick
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2019-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9795680
• 关键词: 3-Dimensional; Address; Affect; Air; Animals; Area; Behavioral; Biological Models; Biomedical Research; Bone Marrow; Cancer Research Project; Carbon Dioxide; Cell Culture System; Cell Culture Techniques; Cell Hypoxia; Cell Line; Cell Survival; Cells; Complement; Core Facility; Data; Development; Disease; Disease model; Elements; Environment; Evaluation; Freezing; Future; Genomics; Goals; Health; Hematopoietic; Histology; Human; Hypoxia; Image; In Situ; In Vitro; Incubators; Inflammation; Injury; Literature; Malignant Neoplasms; Medical; Medical center; Mesenchymal Stem Cells; Methodology; Microfluidic Microchips; Modeling; Molecular; Mus; Myocardial Infarction; Neoplasm Metastasis; Nitrogen; Oxygen; Pathologic; Pathologic Processes; Physiological; Physiological Processes; Physiology; Recovery; Research; Research Personnel; Resources; Services; Sheep; South Carolina; Standardization; Stem cells; Stroke; System; Technology; Time; Tissue Culture Techniques; Tissues; Translations; United States National Institutes of Health; Universities; Veterans; cancer cell; cell injury; cell type; design; experimental study; health care quality; human stem cells; improved; instrument; instrumentation; neoplastic cell; novel; organ on a chip; physiologic model; prevent; programs; response; response to injury; tissue culture; tool; tumor; wasting

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.

该提案的主要目标是建立一个综合低氧细胞核心设施（HCCF），作为 现有的细胞和分子评价核心（CMEC）设施内的研究服务在拉尔夫H。 约翰逊退伍军人医疗中心（RHJVAMC）。HCCF将围绕三个 最先进的HypOxygen低氧工作站，每个都具有新颖的功能， 将缺氧细胞培养箱和处理站集成在一个单元中。这将使长期 在单一受控的氧、CO2和氮环境中传代和处理细胞。 此外，作为一个全方位服务的核心，HCCF将包括一个受控速率细胞冷冻系统， 标准化细胞冷冻，减少变异性，优化加工、冷冻、储存和解冻 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，不仅用于尖端研究，还用于需要低氧细胞培养的关键领域 在RHJVAMC的干细胞，癌症和缺血性损伤研究计划，但这是解决一个 新出现的问题被认为是所有体外和原位研究的障碍，即生理学 用于细胞和组织的处理、培养和研究的氧气环境。现在大家都知道， 标准“常氧”或室内空气组织培养技术是非生理性的， 研究数据的解释。这也满足了VA和NIH最近的紧急呼吁， 适当的生理相关的“正常”和疾病模型系统，以防止浪费资源， 这些实验无法模拟真实的世界生理学，并混淆了研究文献。因此，可用性 低氧细胞核心设施的建立不仅可以进行与细胞和组织病理性减少有关的研究， 在疾病和受伤中看到的氧气，但它将提高VA调查人员执行“标准”组织的能力， 以更生理相关的方式培养，支持和推进直接影响人类健康的研究。 我们退伍军人的健康。我们的研究人员已经确定了四个主要目标，将通过这一点来实现 突破性的技术和集成的核心：1）需要缺氧细胞培养仪器， 长期培养干细胞，这些干细胞通常存在于“缺氧”的微环境中。2)需要 评估病理性低氧在组织/细胞损伤（例如炎症和缺血性损伤，包括 中风和心肌梗塞），并用于肿瘤中发现的不同缺氧区和缺氧区的建模; 3） 允许将现有或未来的CMEC仪器放置在低氧室内，用于活细胞/组织的实时检测， 分析;和4）改善难以冷冻和回收的细胞系的处理、储存和回收， 特别是人类干细胞和原代细胞。至关重要的是，这一核心将填补RHJVAMC的一个主要未满足的需求 因为在VA或其附属机构内没有可识别的缺氧细胞培养系统， 南卡罗来纳州医科大学。最先进的缺氧细胞培养技术， Hypoxygen工作站和其他核心元素，以及HCCF的加入， VA CMEC将大大加强我们站的研究能力，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 快速实施先进的个性化医疗，为我们的客户提供优质的医疗服务， 国家的退伍军人，在部长的卓越蓝图和部长舒尔金的具体指令概述 10-点计划。