Department of Mechanical and Aerospace Engineering, NC State University, Raleigh, USA.
Department of Biomedical Engineering, University of Mississippi, Oxford, MS, USA.
Sci Rep. 2021 Feb 17;11(1):4020. doi: 10.1038/s41598-021-83579-1.
Tumor tissue contains a continuous distribution of static and dynamically changing oxygen environments with levels ranging from physiologically normal oxygen down to anoxia. However, in vitro studies are often performed under oxygen levels that are far higher than those found in vivo. A number of devices are available to alter the oxygen environment in cell culture, including designs from our laboratory. However, in our devices and most other designs, changing the media in order to feed or dose cells remains a disruptive factor in maintaining a consistent hypoxic environment. This report presents a novel 96-well plate design that recirculates the local oxygen environment to shield cells during media changes and facilitates toxicity studies of cells cultured under varying oxygen levels. The principle behind the design is presented and the response of human pancreatic cancer PANC-1 cells treated with tirapazamine and doxorubicin under eight different static or cycling oxygen levels was measured. As expected, tirapazamine is progressively more toxic as oxygen levels decrease but retains some toxicity as oxygen is cycled between hypoxic and normoxic levels. Doxorubicin sensitivity is largely unaffected by changing oxygen levels. This technology is ideal for assessing the effects of oxygen as a variable in toxicity screens.
肿瘤组织包含静态和动态变化的氧气环境的连续分布，氧水平从生理正常氧降至缺氧。然而，体外研究通常在远高于体内发现的氧水平下进行。有许多设备可用于改变细胞培养中的氧气环境，包括我们实验室的设计。然而，在我们的设备和大多数其他设计中，为了喂养或处理细胞而改变培养基仍然是维持一致低氧环境的一个干扰因素。本报告介绍了一种新颖的 96 孔板设计，该设计可在培养基变化期间循环局部氧气环境以保护细胞，并促进在不同氧水平下培养的细胞的毒性研究。介绍了设计背后的原理，并测量了在八种不同静态或循环氧水平下用替拉扎明和阿霉素处理的人胰腺癌细胞 PANC-1 的反应。正如预期的那样，随着氧水平的降低，替拉扎明的毒性逐渐增加，但在缺氧和正常氧之间循环时仍保留一些毒性。改变氧水平对阿霉素的敏感性影响不大。该技术非常适合评估氧气作为毒性筛选中变量的影响。
