ShEEP application for Integrated Hypoxia Exposure and Analysis Core

Ralph H Johnson Va Medical Center, Charleston SC

Investigators

Abstract

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.