Related Experiment Video
Updated: Jun 15, 2026
Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis
Published on: December 23, 2022
Tumor collection/processing under physioxia uncovers highly relevant signaling networks and drug sensitivity
Brijesh Kumar1, Adedeji K Adebayo1,2, Mayuri Prasad1
1Department of Surgery, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Abstract:
Preclinical studies of primary cancer cells are typically done after tumors are removed from patients or animals at ambient atmospheric oxygen (O2, ~21%). However, O2 concentrations in organs are in the ~3 to 10% range, with most tumors in a hypoxic or 1 to 2% O2 environment in vivo. Although effects of O2 tension on tumor cell characteristics in vitro have been studied, these studies are done only after tumors are first collected and processed in ambient air. Similarly, sensitivity of primary cancer cells to anticancer agents is routinely examined at ambient O2. Here, we demonstrate that tumors collected, processed, and propagated at physiologic O2 compared to ambient air display distinct differences in key signaling networks including LGR5/WNT, YAP, and NRF2/KEAP1, nuclear reactive oxygen species, alternative splicing, and sensitivity to targeted therapies. Therefore, evaluating cancer cells under physioxia could more closely recapitulate their physiopathologic status in the in vivo microenvironment.
Insights
Cancer cells studied in physiologic oxygen (physioxia) show different signaling and drug responses compared to those studied in ambient air. This suggests physioxia better reflects the tumor microenvironment for accurate preclinical research.
Area of Science:
- Oncology
- Cell Biology
- Biomedical Research
Background:
- Preclinical cancer studies often use ambient oxygen (~21%), not the lower oxygen levels found in tumors.
- Tumor cells in vivo typically exist in a hypoxic environment (1-10% oxygen).
- Current in vitro studies process tumors in ambient air, potentially altering cell characteristics.
Purpose of the Study:
- To investigate the impact of oxygen levels on primary cancer cell characteristics.
- To compare cancer cell behavior and drug sensitivity under physiologic oxygen versus ambient air.
- To determine if studying cancer cells under physioxia better mimics their in vivo state.
Main Methods:
- Collected, processed, and propagated primary tumors under controlled physiologic oxygen conditions (physioxia).
- Compared key cellular signaling networks (LGR5/WNT, YAP, NRF2/KEAP1) between physioxia and ambient air groups.
- Assessed differences in nuclear reactive oxygen species, alternative splicing, and sensitivity to targeted therapies.
Main Results:
- Cancer cells cultured under physioxia exhibited distinct differences in signaling networks compared to ambient air.
- Physiologic oxygen conditions altered nuclear reactive oxygen species and alternative splicing patterns.
- Sensitivity to targeted anticancer therapies varied significantly based on oxygen exposure during processing and propagation.
Conclusions:
- Processing and culturing cancer cells under physiologic oxygen (physioxia) significantly impacts their molecular profiles and drug responses.
- Current standard methods using ambient air may not accurately represent the in vivo tumor microenvironment.
- Evaluating cancer cells under physioxia is crucial for more relevant preclinical research and drug development.
