Zdrazilova 2021 MitoFit ace-sce: Difference between revisions
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|title=Zdrazilova L, | |title=Zdrazilova L, Hansikova H, Gnaiger E (2021) Comparable respiratory activity in attached and suspended human fibroblasts. https://doi.org/10.26124/mitofit:2021-0007 β ''2022-03-03 published in [[Zdrazilova 2022 PLOS ONE |PLOS ONE 17:e0264496]]'' | ||
|info=in | |info=MitoFit Preprints 2021.07. [[File:MitoFit Preprints pdf.png|left|160px|link=https://www.mitofit.org/images/1/15/Zdrazilova_2021_MitoFit_ace-sce.pdf |MitoFit pdf]] [https://www.mitofit.org/images/1/15/Zdrazilova_2021_MitoFit_ace-sce.pdf Comparable respiratory activity in attached and suspended human fibroblasts]<br/> | ||
|authors= | |authors=MitoFit Prep 2021.7. | ||
|year=2021 | |year=2021 | ||
|journal=MitoFit | |journal=MitoFit Prep | ||
|abstract=Measurement of oxygen consumption of cultured cells is widely used for diagnosis of mitochondrial diseases, drug testing, biotechnology and toxicology. Fibroblasts are cultured in monolayers but physiological measurements are carried out in suspended or attached cells. We address the question whether respiration differs in attached and suspended cells using multiwell respirometry (Agilent Seahorse XF24) and high-resolution respirometry (Oroboros O2k), respectively. Respiration of human dermal fibroblasts measured in culture medium was baseline-corrected for residual oxygen consumption and expressed as oxygen flow per cell. | |abstract='''Journal publication 2022-03-03 in [[Zdrazilova 2022 PLOS ONE |Β»'''PLOS ONE 17:e0264496'''Β«]]''' | ||
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Version 1 ('''v1''') '''2021-09-21''' [https://www.mitofit.org/images/1/15/Zdrazilova_2021_MitoFit_ace-sce.pdf doi:10.26124/mitofit:2021-0007] | |||
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Measurement of oxygen consumption of cultured cells is widely used for diagnosis of mitochondrial diseases, drug testing, biotechnology and toxicology. Fibroblasts are cultured in monolayers but physiological measurements are carried out in suspended or attached cells. We address the question whether respiration differs in attached and suspended cells using multiwell respirometry (Agilent Seahorse XF24) and high-resolution respirometry (Oroboros O2k), respectively. Respiration of human dermal fibroblasts measured in culture medium was baseline-corrected for residual oxygen consumption and expressed as oxygen flow per cell. | |||
No differences were observed in ROUTINE respiration of living cells and LEAK respiration obtained after inhibition of ATP synthase by oligomycin. Multiple steps of uncoupler titrations in the O2k allowed for evaluation of maximum electron transfer capacity, which was higher than respiration obtained in the XF24 due to a limitation to two uncoupler titrations. | No differences were observed in ROUTINE respiration of living cells and LEAK respiration obtained after inhibition of ATP synthase by oligomycin. Multiple steps of uncoupler titrations in the O2k allowed for evaluation of maximum electron transfer capacity, which was higher than respiration obtained in the XF24 due to a limitation to two uncoupler titrations. | ||
Quantitative evaluation of respiration measured in different platforms revealed that short-term suspension of fibroblasts did not affect respiratory activity and coupling control. Consistent results obtained with different platforms provide a test for reproducibility and allow for building an extended respirometric database. | Quantitative evaluation of respiration measured in different platforms revealed that short-term suspension of fibroblasts did not affect respiratory activity and coupling control. Consistent results obtained with different platforms provide a test for reproducibility and allow for building an extended respirometric database. | ||
|keywords=attached cells ace, suspended cells sce, fibroblasts, high-resolution respirometry HRR, Oroboros O2k, Seahorse Bioanalyzer XF24, ROUTINE respiration ''R'', LEAK respiration ''L'', electron transfer capacity ''E'', residual oxygen consumption ''Rox'', bioenergetic cluster analysis BCA | <br><br> | ||
|keywords=attached cells; ace, suspended cells; sce, fibroblasts, high-resolution respirometry; HRR, Oroboros O2k, Seahorse Bioanalyzer XF24, ROUTINE respiration; ''R'', LEAK respiration; ''L'', electron transfer capacity; ''E'', residual oxygen consumption; ''Rox'', bioenergetic cluster analysis; BCA | |||
|editor=Gnaiger E | |editor=Gnaiger E | ||
|mipnetlab=AT Innsbruck Oroboros | |mipnetlab=AT Innsbruck Oroboros, CZ Prague Zeman J | ||
}} | }} | ||
ORC'''ID''': [[File:ORCID.png|20px|link=https://orcid.org/0000-0002-8103-2498]] Zdrazilova Lucie, [[File:ORCID.png|20px|link=https://orcid.org/0000-0002-2734-225X]] Hanskova Hana, [[File:ORCID.png|20px|link=https://orcid.org/0000-0003-3647-5895]] Gnaiger Erich | |||
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== Data availability == | |||
:::: Original files are available Open Access at Zenodo repository: https://zenodo.org/record/5518059#.YUmBqn2xWCh | |||
== Support == | == Support == | ||
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== Cited by == | == Cited by == | ||
{{Template:Cited by Gnaiger 2021 MitoFit BCA}} | {{Template:Cited by Gnaiger 2021 MitoFit BCA}} | ||
{{Template:Cited by Krako Jakovljevic 2021 BEC PD}} | |||
== Reference to an Abstract == | |||
::::* Felser A, Larsson NG (2014) Assessing mitochondrial dysfunction in fibroblast cells β a comparison between O2k and multiwell respirometry. Mitochondr Physiol Network 19.13 - MiP2014. - [[Felser 2014 Abstract MiP2014 |Β»Bioblast linkΒ«]] | |||
{{Labeling | {{Labeling | ||
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Latest revision as of 07:16, 3 October 2023
Zdrazilova 2021 MitoFit ace-sce
Zdrazilova L, Hansikova H, Gnaiger E (2021) Comparable respiratory activity in attached and suspended human fibroblasts. https://doi.org/10.26124/mitofit:2021-0007 β 2022-03-03 published in PLOS ONE 17:e0264496 |
Β» MitoFit Preprints 2021.07.
Comparable respiratory activity in attached and suspended human fibroblasts
MitoFit Prep 2021.7. (2021) MitoFit Prep
Abstract: Journal publication 2022-03-03 in Β»PLOS ONE 17:e0264496Β«
Version 1 (v1) 2021-09-21 doi:10.26124/mitofit:2021-0007
Measurement of oxygen consumption of cultured cells is widely used for diagnosis of mitochondrial diseases, drug testing, biotechnology and toxicology. Fibroblasts are cultured in monolayers but physiological measurements are carried out in suspended or attached cells. We address the question whether respiration differs in attached and suspended cells using multiwell respirometry (Agilent Seahorse XF24) and high-resolution respirometry (Oroboros O2k), respectively. Respiration of human dermal fibroblasts measured in culture medium was baseline-corrected for residual oxygen consumption and expressed as oxygen flow per cell.
No differences were observed in ROUTINE respiration of living cells and LEAK respiration obtained after inhibition of ATP synthase by oligomycin. Multiple steps of uncoupler titrations in the O2k allowed for evaluation of maximum electron transfer capacity, which was higher than respiration obtained in the XF24 due to a limitation to two uncoupler titrations.
Quantitative evaluation of respiration measured in different platforms revealed that short-term suspension of fibroblasts did not affect respiratory activity and coupling control. Consistent results obtained with different platforms provide a test for reproducibility and allow for building an extended respirometric database.
β’ Keywords: attached cells; ace, suspended cells; sce, fibroblasts, high-resolution respirometry; HRR, Oroboros O2k, Seahorse Bioanalyzer XF24, ROUTINE respiration; R, LEAK respiration; L, electron transfer capacity; E, residual oxygen consumption; Rox, bioenergetic cluster analysis; BCA
β’ Bioblast editor: Gnaiger E
β’ O2k-Network Lab: AT Innsbruck Oroboros, CZ Prague Zeman J
ORCID: Zdrazilova Lucie, Hanskova Hana, Gnaiger Erich
Data availability
- Original files are available Open Access at Zenodo repository: https://zenodo.org/record/5518059#.YUmBqn2xWCh
Support
- This work was partially funded by the European Unionβs Horizon 2020 research and innovation programme under grant agreement No. 859770, NextGen-O2k project, and by Institutional project GAUK110119. Contribution to COST Action CA15203 MitoEAGLE with financial support of Short-Term Scientific missions (LD).
Cited by
- Gnaiger E (2021) Bioenergetic cluster analysis β mitochondrial respiratory control in human fibroblasts. MitoFit Preprints 2021.8. https://doi.org/10.26124/mitofit:2021-0008
- Krako Jakovljevic N, Ebanks B, Katyal G, Chakrabarti L, Markovic I, Moisoi N (2021) Mitochondrial homeostasis in cellular models of Parkinsonβs Disease. Bioenerg Commun 2021.2. https://doi.org/10.26124/bec:2021-0002
- Gnaiger E (2021) Bioenergetic cluster analysis β mitochondrial respiratory control in human fibroblasts. MitoFit Preprints 2021.8. https://doi.org/10.26124/mitofit:2021-0008
Reference to an Abstract
- Felser A, Larsson NG (2014) Assessing mitochondrial dysfunction in fibroblast cells β a comparison between O2k and multiwell respirometry. Mitochondr Physiol Network 19.13 - MiP2014. - Β»Bioblast linkΒ«
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