Difference between revisions of "Zhang 2012 Nat Protoc"

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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. • Keywords: OROBOROS vs. Seahorse

Labels:

HRR: Oxygraph-2k

Further information

* Also see: Mitochondrial preparations - Cultured cells

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 {{Publication
 |title=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.
-|info=[http://www.ncbi.nlm.nih.gov/pubmed?term=Measuring%20energy%20metabolism%20in%20cultured%20cells%2C%20including%20human%20pluripotent%20stem%20cells%20and%20differentiated%20cells PMID: 22576106]
+|info=[http://www.ncbi.nlm.nih.gov/pubmed?term=Measuring%20energy%20metabolism%20in%20cultured%20cells%2C%20including%20human%20pluripotent%20stem%20cells%20and%20differentiated%20cells PMID: 22576106], [http://www.nature.com/nprot/journal/v7/n6/fig_tab/nprot.2012.048_T1.html Table: Comparison of common bioenergetic analysis methods]
 |authors=Zhang J, Nuebel E, Wisidagama DR, Setoguchi K, Hong JS, Van Horn CM, Imam SS, Vergnes L, Malone CS, Koehler CM, Teitell MA
 |year=2012
@@ Line 11: / Line 11: @@
 |instruments=Oxygraph-2k
 }}
-* [http://www.nature.com/nprot/journal/v7/n6/fig_tab/nprot.2012.048_T1.html Comparison of common bioenergetic analysis methods]
+== Further information ==
-:: * '''The O2k in cell culture apllications''':
-::* Pesta D, Gnaiger E (2012) [[Pesta 2012 Methods Mol Biol|High-resolution respirometry. OXPHOS  protocols for human cells and permeabilized fibres from small biopisies  of human muscle.]] Methods Mol Biol 810: 25-58.
-::* Garedew A, Haffner B, Huetter E, Gnaiger E.[[MiPNet10.04 CellRespiration|An experiment with  high-resolution respirometry: Phosphorylation control in cell  respiration.]] Mitochondr Physiol Network 10.04.
-::* Gnaiger E.[[MiPNet11.05 Mitos-PermeabilizedCells|Isolated mitochondria or permeabilized tissues and cells.]] Mitochondr Physiol Network 11.05.
 :: * Also see: [[O2k-Publications: Intact Cell; Cultured; Primary|Mitochondrial preparations - Cultured cells]]