Martinez Chloe-Anne, Cistulli Peter A, Cook Kristina M
The University of Sydney, Charles Perkins Centre, Northern Clinical School, New South Wales, Australia.
Bio Protoc. 2019 Sep 20;9(18):e3371. doi: 10.21769/BioProtoc.3371.
Dissolved oxygen and its availability to cells in culture is an overlooked variable which can have significant consequences on experimental research outcomes, including reproducibility. Oxygen sensing pathways play key roles in cell growth and behavior and pericellular oxygen levels should be controlled when establishing models. Standard cell culture techniques do not have adequate control over pericellular oxygen levels. Slow diffusion through culture media limits the precision of oxygen delivery to cells, making it difficult to accurately reproduce -like oxygen conditions. Furthermore, different types of cells consume oxygen at varying rates and this can be affected by the density of growing cells. Here, we describe a novel system that utilizes hypoxic chambers and oxygen-permeable culture dishes to control pericellular oxygen levels and provide rapid oxygen delivery to adherent cells. This procedure is particularly relevant for protocols studying effects of rapid oxygen changes or intermittent hypoxia on cellular behavior. The system is inexpensive and easily assembled without highly specialized equipment.
溶解氧及其在培养细胞中的可利用性是一个被忽视的变量，它可能对实验研究结果产生重大影响，包括可重复性。氧传感途径在细胞生长和行为中起关键作用，在建立模型时应控制细胞周围的氧水平。标准细胞培养技术对细胞周围氧水平的控制不足。通过培养基的缓慢扩散限制了向细胞输送氧气的精度，使得难以准确再现类似的氧气条件。此外，不同类型的细胞以不同的速率消耗氧气，这可能会受到生长细胞密度的影响。在这里，我们描述了一种新颖的系统，该系统利用缺氧箱和透氧培养皿来控制细胞周围的氧水平，并为贴壁细胞提供快速的氧气输送。该程序对于研究快速氧气变化或间歇性缺氧对细胞行为影响的实验方案尤为重要。该系统价格低廉，无需高度专业化的设备即可轻松组装。
