Related Experiment Video
Updated: Oct 1, 2025
10:26
Assessment of Mitochondrial Health in Cancer-Associated Fibroblasts Isolated from 3D Multicellular Lung Tumor Spheroids
Published on: October 21, 2022
2.1K
Comparable respiratory activity in attached and suspended human fibroblasts
Lucie Zdrazilova1, Hana Hansikova1, Erich Gnaiger2
1Department of Pediatrics and Inherited Metabolic Disorders, First Faculty of Medicine, Charles University and General Hospital in Prague, Prague, Czechia.
Plos One
|March 3, 2022
Summary
Cellular respiration measurements are consistent whether cells are attached or suspended. This finding supports the reliability of respirometry data for mitochondrial disease diagnosis and drug testing.
Area of Science:
- Cellular respiration and mitochondrial function analysis.
- Biotechnology and toxicological applications of respirometry.
Background:
- Oxygen consumption measurement is crucial for diagnosing mitochondrial diseases, drug testing, and toxicology.
- Fibroblast cell cultures are typically grown as monolayers, but physiological measurements often involve suspended or attached cells.
Purpose of the Study:
- To investigate whether cellular respiration differs between attached and suspended human dermal fibroblasts.
- To compare results from multiwell respirometry (Agilent Seahorse XF24) and high-resolution respirometry (Oroboros O2k).
Main Methods:
- Human dermal fibroblasts were cultured and their oxygen consumption measured using Agilent Seahorse XF24 (attached cells) and Oroboros O2k (suspended cells).
- Respiration measurements included ROUTINE and LEAK respiration (after oligomycin inhibition).
- Baseline correction for residual oxygen consumption was performed, and results were expressed as oxygen flow per cell.
Main Results:
- No significant differences were observed in ROUTINE or LEAK respiration between attached and suspended fibroblasts.
- Electron transfer capacity measurements were higher with the Oroboros O2k compared to the Agilent Seahorse XF24, potentially due to titration differences.
- Short-term suspension did not impact fibroblast respiratory activity or coupling control.
Conclusions:
- Cellular respiration measurements in fibroblasts are reliable regardless of whether cells are attached or suspended.
- Differences in electron transfer capacity measurements between platforms highlight the importance of standardized titration protocols.
- Validating respirometric data across different platforms is essential for building a reliable database for research and diagnostics.
