Oxygen-Regulated Tissue Culturing for Accurate In Virto Cell Analyses

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We request a gas-controlled cell culturing station that allows the establishment and culturing of cells under conditions of oxygen that are similar to what cells encounter in the body (normoxia). Culturing of cells under ambient (20%) oxygen concentrations that is usually done causes many cells to rapidly stop growing due to the oxidative stress that impacts many growth and repair pathways. In contrast, culturing cells in 2-5% oxygen has been shown to significantly extend the replicative lifespan of mouse cells from 5-10 cell divisions to >60, to increase their growth rate and to alter DNA repair pathways, showing that oxidative stress strongly impacts cells once they are removed from the body. Members of our group rely heavily upon cell culturing in vitro for our basic research programs. Although our group has two small oxygen-regulated incubators that partly address this problem of chronic oxidative stress, we do not have the tissue culturing facilities needed for proper establishment and culturing of cells that are needed to accurately examine the induction and repair of DNA damage, the regulation of cell replicative lifespan in different species and the factors involved in regulating the formation and activity of membraneless organelles. Without the ability to rigorously control oxygen levels in the absence of the oxygen spikes that are encountered in standard culturing hoods, the results of in vitro experiments do not represent conditions in vivo. This tissue culturing facility will address this shortcoming, be the only one at our university and would provide the members of our group, and the 48 trainees and personnel in our seven labs a unique ability to advance their training using a state-of-the-art culturing facility. Otherwise unattainable data in areas of cell aging, DNA repair and membraneless organelle formation and function will promote the careers of our developing pool of HQP and promote expansion of our training program in our institution and in other labs in Canada that we collaborate with.