Difference between revisions of "Zhang 2012 Nat Protoc"
| Line 6: | Line 6: | ||
|journal=Nat Protoc | |journal=Nat Protoc | ||
|abstract=Measurements of glycolysis and mitochondrial function are required to quantify energy metabolism in a wide variety of cellular contexts. In human pluripotent stem cells (hPSCs) and their differentiated progeny, this analysis can be challenging because of the unique cell properties, growth conditions and expense required to maintain these cell types. Here we provide protocols for analyzing energy metabolism in hPSCs and their early differentiated progenies that are generally applicable to mature cell types as well. Our approach has revealed distinct energy metabolism profiles used by hPSCs, differentiated cells, a variety of cancer cells and Rho-null cells. The protocols measure or estimate glycolysis on the basis of the extracellular acidification rate, and they measure or estimate oxidative phosphorylation on the basis of the oxygen consumption rate. Assays typically require 3 h after overnight sample preparation. Companion methods are also discussed and provided to aid researchers in developing more sophisticated experimental regimens for extended analyses of cellular bioenergetics. | |abstract=Measurements of glycolysis and mitochondrial function are required to quantify energy metabolism in a wide variety of cellular contexts. In human pluripotent stem cells (hPSCs) and their differentiated progeny, this analysis can be challenging because of the unique cell properties, growth conditions and expense required to maintain these cell types. Here we provide protocols for analyzing energy metabolism in hPSCs and their early differentiated progenies that are generally applicable to mature cell types as well. Our approach has revealed distinct energy metabolism profiles used by hPSCs, differentiated cells, a variety of cancer cells and Rho-null cells. The protocols measure or estimate glycolysis on the basis of the extracellular acidification rate, and they measure or estimate oxidative phosphorylation on the basis of the oxygen consumption rate. Assays typically require 3 h after overnight sample preparation. Companion methods are also discussed and provided to aid researchers in developing more sophisticated experimental regimens for extended analyses of cellular bioenergetics. | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
| | |additional=Respiration | ||
}} | }} | ||
::>> [http://www.nature.com/nprot/journal/v7/n6/fig_tab/nprot.2012.048_T1.html Table: Comparison of common bioenergetic analysis methods] | |||
Continue the discussion: [[O2k_versus_multiwell_respirometer]] | Continue the discussion: [[O2k_versus_multiwell_respirometer]] | ||
::* See also: [[Horan 2012 J Gerontol A Biol Sci Med Sci|Review: Quantifying mitochondrial dysfunction in complex diseases of aging (Horan et al 2012)]] | ::* See also: [[Horan 2012 J Gerontol A Biol Sci Med Sci|Review: Quantifying mitochondrial dysfunction in complex diseases of aging (Horan et al 2012)]] | ||
::* Discussion: [[Talk:Rogers 2011 PLoS One| | ::* Discussion: [[Talk:Rogers 2011 PLoS One|XFe for isolated mitochondria - high throughput without high output?]] | ||
* Further information: [[O2k-Publications: Intact Cell; Cultured; Primary]] | * Further information: [[O2k-Publications: Intact Cell; Cultured; Primary]] | ||
Revision as of 10:42, 26 January 2013
| Zhang J, Nuebel E, Wisidagama DR, Setoguchi K, Hong JS, Van Horn CM, Imam SS, Vergnes L, Malone CS, Koehler CM, Teitell MA (2012) Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells. Nat Protoc 7: 1068-1085. |
» PMID: 22576106, Table: Comparison of common bioenergetic analysis methods
Zhang J, Nuebel E, Wisidagama DR, Setoguchi K, Hong JS, Van Horn CM, Imam SS, Vergnes L, Malone CS, Koehler CM, Teitell MA (2012) Nat Protoc
Abstract: Measurements of glycolysis and mitochondrial function are required to quantify energy metabolism in a wide variety of cellular contexts. In human pluripotent stem cells (hPSCs) and their differentiated progeny, this analysis can be challenging because of the unique cell properties, growth conditions and expense required to maintain these cell types. Here we provide protocols for analyzing energy metabolism in hPSCs and their early differentiated progenies that are generally applicable to mature cell types as well. Our approach has revealed distinct energy metabolism profiles used by hPSCs, differentiated cells, a variety of cancer cells and Rho-null cells. The protocols measure or estimate glycolysis on the basis of the extracellular acidification rate, and they measure or estimate oxidative phosphorylation on the basis of the oxygen consumption rate. Assays typically require 3 h after overnight sample preparation. Companion methods are also discussed and provided to aid researchers in developing more sophisticated experimental regimens for extended analyses of cellular bioenergetics.
Labels:
Respiration
Continue the discussion: O2k_versus_multiwell_respirometer
- Further information: O2k-Publications: Intact Cell; Cultured; Primary
