Controlling pericellular oxygen tension in cell culture reveals distinct breast cancer responses to low oxygen tensions
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ABSTRACT:
PROVIDER: PRJNA1129053 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR29620611_1.fastq.gz | Fastqsanger.gz | |||
| SRR29620611_2.fastq.gz | Fastqsanger.gz | |||
| SRR29620612_1.fastq.gz | Fastqsanger.gz | |||
| SRR29620612_2.fastq.gz | Fastqsanger.gz | |||
| SRR29620613_1.fastq.gz | Fastqsanger.gz |
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Project description:In this study, the relationship between pericellular and gas phase oxygen concentration in cell culture was examined. To this end, we examined the influence of cellular oxygen consumption on primary human hepatocyte culture in physioxic (6% O2) and normoxic (18.6% O2) conditions. To understand how pericellular oxygen tension influenced hepatocyte biology in in vitro cell culture, we performed RNA-seq (this dataset).
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Project description:In oxygen (O2)-controlled cell culture, an indispensable tool in biological research, it is presumed that the incubator setpoint equals the O2 tension experienced by cells (i.e., pericellular O2). However, it is discovered that physioxic (5% O2) and hypoxic (1% O2) setpoints regularly induce anoxic (0% O2) pericellular tensions in both adherent and suspension cell cultures. Electron transport chain inhibition ablates this effect, indicating that cellular O2 consumption is the driving factor. RNA-seq analysis revealed that primary human hepatocytes cultured in physioxia experience ischemia-reperfusion injury due to cellular O2 consumption. A reaction-diffusion model is developed to predict pericellular O2 tension a priori, demonstrating that the effect of cellular O2 consumption has the greatest impact in smaller volume culture vessels. By controlling pericellular O2 tension in cell culture, it is found that hypoxia vs. anoxia induce distinct breast cancer transcriptomic and translational responses, including modulation of the hypoxia-inducible factor (HIF) pathway and metabolic reprogramming. Collectively, these findings indicate that breast cancer cells respond non-monotonically to low O2, suggesting that anoxic cell culture is not suitable for modeling hypoxia. Furthermore, it is shown that controlling atmospheric O2 tension in cell culture incubators is insufficient to regulate O2 in cell culture, thus introducing the concept of pericellular O2-controlled cell culture.
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