SUIT-003 O2 ce-pce D020: Difference between revisions
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{{MitoPedia | {{MitoPedia | ||
|abbr=ceP,S | |abbr=ceP,S | ||
|description=[[File:ce1;ce2P;ce3Omy;ce4U;ce5Rot;ce6S;1Dig;1c;2Ama;3AsTm;4Azd.png|400px]] | |description=[[File:ce1;ce2P;ce3Omy;ce4U;ce5Rot;ce6S;1Dig;1c;2Ama;3AsTm;4Azd.png|400px]][[File:D020 O2 traces.png|400px]] | ||
|info='''A''' '''[[SUIT-003]]''' Coupling control protocol and respirometric cell viability test '''(CCV protocol)''' Β | |info='''A''' '''[[SUIT-003]]''' Coupling control protocol and respirometric cell viability test '''(CCV protocol)''' Β | ||
|application=O2 | |application=O2 |
Revision as of 14:36, 28 February 2019
Description
Abbreviation: ceP,S
Reference: A SUIT-003 Coupling control protocol and respirometric cell viability test (CCV protocol)
SUIT number: D020_ce1;ce2P;ce3Omy;ce4U;ce5Rot;ce6S;1Dig;1c;2Ama;3AsTm;4Azd
O2k-Application: O2
- SUIT-category: ceP,S
- SUIT protocol pattern: complex cell coupling control protocol ce1;ce2P;ce3Omy;ce4U;ce5Rot;ce6S;1Dig;1c;2Ama;3AsTm;4Azd
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce1P | ROUTINE | ce1;ce1P
| ||
ce2Omy | L(Omy) | ce1;ce1P;ce2Omy
| ||
ce3U | E | ce1;ce1P;ce2Omy;ce3U
| ||
ce4Rot | ROX | ce1;ce1P;ce2Omy;ce3U;ce4Rot
|
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce5S | ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S
| |||
1Dig* | SE | S | CII | ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;1Dig
|
1c | SE | S | CII | ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;1Dig;1c
|
2Ama | ROX | ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;1Dig;1c;2Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
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- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- The SUIT-003 O2 ce-pce D020 is a cell coupling control protocol. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce).
- For determination of respirometric viability index, it is assumed that both viable and dead cells contain functional mitochondria.
- When the cell viability test module is not incorporated into the experiment, complex III inhibitors can be used to reach Rox instead of rotenone.
- + Respirometric cell viability test correlates with other cell viability quantification methods.
- + With the extended module, S-ET capacity can be assessed after the performance of the coupling control protocol, as well as outer mt-membrane integrity with the addition of cytochrome c.
- + CIV activity can also be measured after the cell viability test module.
- + Flexibility of experiment duration: from 1 hour (cell coupling control protocol) to 2 and a half hours (cell coupling control, cell viability test module and CIV module).
- - Cells with a cell membrane succinate transporter are not suitable for the viability test.
- - Pathological profiles could hinder the cell viability test.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
- - The addition of oligomycin has to be carefully tested to avoid inhibitory effects in the evaluation of the ET-capacity.
Compare SUIT protocols
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
Doerrier 2018 Methods Mol Biol | 2018 | Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, MΓ©szΓ‘ros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70. https://doi.org/10.1007/978-1-4939-7831-1_3 | Human Mouse Rat Saccharomyces cerevisiae | Heart Skeletal muscle Endothelial;epithelial;mesothelial cell Blood cells HEK Platelet |
Stadlmann 2006 Cell Biochem Biophys | 2006 | Stadlmann S, Renner K, Pollheimer J, Moser PL, Zeimet AG, Offner FA, Gnaiger E (2006) Preserved coupling of oxidative phosphorylation but decreased mitochondrial respiratory capacity in IL-1Γ treated human peritoneal mesothelial cells. Cell Biochem Biophys 44:179-86. | Human | Endothelial;epithelial;mesothelial cell |
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry