Difference between revisions of "Zdrazilova 2021 MitoFit ace-sce"
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{{Publication | {{Publication | ||
|title=Zdrazilova L, Hanskova H, Gnaiger E (2021) Comparable respiratory activity in attached and suspended human fibroblast cell lines. MitoFit Preprints 2021.#. [[doi:10.26124/mitofit:2021-000#]] (in preparation) | |title=Zdrazilova L, Hanskova H, Gnaiger E (2021) Comparable respiratory activity in attached and suspended human fibroblast cell lines. MitoFit Preprints 2021.#. [[doi:10.26124/mitofit:2021-000#]] (in preparation) | ||
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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 | |||
|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 | ||
}} | }} | ||
{{Labeling | |||
|area=Respiration, Instruments;methods | |||
|organism=Human | |||
|tissues=Fibroblast | |||
|preparations=Intact cells | |||
|topics=Coupling efficiency;uncoupling | |||
|couplingstates=LEAK, OXPHOS, ET | |||
|pathways=ROX | |||
|instruments=Oxygraph-2k | |||
|additional=SUIT-003, MitoEAGLEPublication, MitoFit 2021 BCA | |||
}} | |||
{{MitoFit page name}} | |||
ORC'''ID''': Zdrazilova Lucie, Hanskova Hana, [[File:ORCID.png|20px|link=https://orcid.org/0000-0003-3647-5895]] Gnaiger Erich | ORC'''ID''': Zdrazilova Lucie, Hanskova Hana, [[File:ORCID.png|20px|link=https://orcid.org/0000-0003-3647-5895]] Gnaiger Erich | ||
= References = | = References = | ||
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:::: This work was partially funded by the European Union’s 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). | :::: This work was partially funded by the European Union’s 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 == | |||
{{Template:Cited by Gnaiger 2021 MitoFit BCA}} | |||
}} | |||
Revision as of 11:48, 25 August 2021
| Zdrazilova L, Hanskova H, Gnaiger E (2021) Comparable respiratory activity in attached and suspended human fibroblast cell lines. MitoFit Preprints 2021.#. doi:10.26124/mitofit:2021-000# (in preparation) |
» in preparation
Zdrazilova Lucie, Hanskova Hana, Gnaiger Erich (2021-##-##) MitoFit Preprints
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.
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. • 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 • Bioblast editor: Gnaiger E • O2k-Network Lab: AT Innsbruck Oroboros
Labels: MiParea: Respiration, Instruments;methods
Organism: Human
Tissue;cell: Fibroblast
Preparation: Intact cells
Regulation: Coupling efficiency;uncoupling Coupling state: LEAK, OXPHOS, ET Pathway: ROX HRR: Oxygraph-2k
SUIT-003, MitoEAGLEPublication, MitoFit 2021 BCA
Zdrazilova 2021 MitoFit ace-sce
ORCID: Zdrazilova Lucie, Hanskova Hana, 
Gnaiger Erich
References
Support
- This work was partially funded by the European Union’s 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 – mitochondrial respiratory control in human fibroblasts. MitoFit Preprints 2021.8. https://doi.org/10.26124/mitofit:2021-0008
