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.

该提案的主要目的是建立一个综合的缺氧细胞核心设施（HCCF）作为一部分 Ralph H的研究服务中的现有细胞和分子评估核心（CMEC）设施。 约翰逊VA医疗中心（RHJVAMC）。 HCCF将集中在三个 最先进的低氧缺氧工作站，每个工作站都具有新颖的功能 一个单元中的综合缺氧细胞培养孵化器和加工站。这将允许长期 单个受控氧，二氧化碳和氮环境中细胞的传代和处理。 此外，作为全方位服务的核心，HCCF将包括受控的速率单元格系统 标准化细胞冷冻，降低变异性并优化处理，冷冻，存储和解冻 恢复有价值的鼠和人类细胞。该核心设施的总体目标是提供 不仅需要对需要缺氧细胞培养至关重要的领域进行尖端研究必要的工具和服务 对于rhjvamc的干细胞，癌症和缺血性损伤研究计划，但正在解决 新兴问题被认为是所有体外和原位研究的障碍，即身体上 用于处理，培养和研究细胞和组织的氧气环境。现在明白 标准的“常规氧”或房间空气组织培养技术是非生理学的，并在 研究数据的解释。这也符合最近和紧急的VA和NIH呼吁开发 适当相关的“正常”和疾病模型系统，以防止浪费资源 无法模拟现实世界生理学和泥泞研究文献的实验。因此，可用性 缺氧细胞核心设施不仅允许与细胞和组织的病理减少有关的研究 氧气在疾病和损伤中看到，但它将增强VA研究人员执行“标准”组织的能力 以更加相关的方式进行文化，并支持和推进直接影响的研究 我们的退伍军人的健康。我们的调查人员已经确定了四个主要目标 开创性技术和综合核心：1）需要缺氧细胞文化仪器以允许 通常存在于“低氧”小丁基微环境中的干细胞的长期培养。 2）需要 进入组织/细胞损伤中病理低氧的影响（例如感染和缺血性损伤 中风和心肌梗塞），以及用于在肿瘤中发现的不同缺氧和缺氧区域的建模； 3） 允许现有或将来的CMEC仪器放置在实时电池/组织的低氧室内 分析; 4）改善了处理，存储和恢复难以冻结和恢复细胞系的困难和恢复 特定的人类干细胞和原代细胞。至关重要的是，该核心将满足Rhjvamc的主要未满足需求 因为在VA或其关联公司内没有可识别的缺氧细胞聚类系统， 南卡罗来纳州医科大学。最先进的缺氧细胞培养技术由 低氧工作站和其他核心元素以及将HCCF添加到已建立的 VA CMEC将极大地增强我们车站的研究能力，从而促进基础科学发现 物理和病理过程的基础机制，以及这些发现的翻译以支持 快速实施最先进的个性化医疗，以改善我们的质量医疗保健 国家退伍军人，秘书蓝图中概述的特定指令和秘书舒尔金的 10点计划。