Talk:Zhang 2012 Nat Protoc
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Revision as of 19:16, 2 July 2012 by Meissner Barbara (talk | contribs)
Discussion on limitations of this study:
- The authors use a ROCK inhibitor to prevent cells from clumping, but there is no information on whether a possible effect of this inhibitor on mitochondrial respiration has been tested.
- Short periods of high oxygen concentrations can influence stem cell properties. How did the authors transport the stem cells from the incubator to the respirometer - is there hypoxic atmosphere in the respirometer?
- The authors nicely discuss the classical view on cell permeabilization and measurement of Citrate Synthase/Lactate Dehydrogenase, but do not mention the latest publications. E.g. nowadays titration schemes are much simpler.
- 50µM ADP was used. In isolated rat liver mitochondria the apparent Km is 52µM, corresponding to 50% of OXPHOS capacity. The required ADP concentration needed to evaluate Vmax should be minimum 9 times the Km.
