Oxygen-Regulated Tissue Culturing for Accurate In Virto Cell Analyses

我们要求一个气体控制的细胞培养站，允许在类似于细胞在体内遇到的氧气条件下建立和培养细胞(常氧)。在环境(20%)氧浓度下培养细胞，通常会导致许多细胞由于影响许多生长和修复途径的氧化应激而迅速停止生长。相比之下，在2-5%的氧气中培养细胞被证明可以显著延长小鼠细胞的复制寿命，从5-10个细胞分裂到&gt；60，提高它们的生长速度和改变DNA修复途径，表明一旦细胞被移出体内，氧化应激就会对细胞产生强烈的影响。我们团队的成员在很大程度上依赖于体外细胞培养来支持我们的基础研究项目。虽然我们的团队有两个小型的氧气调节孵化器，部分解决了这个慢性氧化应激问题，但我们没有适当建立和培养细胞所需的组织培养设施，这是准确检测DNA损伤的诱导和修复、不同物种中细胞复制寿命的调节以及调节无膜细胞器的形成和活动所涉及的因素所需的。在没有标准培养罩中遇到的氧气尖峰的情况下，如果没有严格控制氧气水平的能力，体外实验的结果就不能代表体内的条件。这个组织培养设施将解决这一缺点，是我们大学唯一的一个，将为我们团队的成员以及我们七个实验室的48名学员和人员提供一种独特的能力，利用最先进的培养设施推进他们的培训。否则，在细胞老化、DNA修复和无膜细胞器形成和功能领域无法获得的数据将促进我们正在开发的HQP池的职业生涯，并促进我们的培训计划在我们机构和与我们合作的加拿大其他实验室的扩展。

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.