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Talk:Zhang 2012 Nat Protoc

From Bioblast

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 transfer the stem cells from the incubator to the respirometer - was there a hypoxic atmosphere in the respirometer?
  • The authors discuss the classical view on cell permeabilization and measurement of citrate synthase/lactate dehydrogenase, but do not mention the latest publications, e.g. up-to-date simple titration schemes.
  • 50 ยตM ADP was used. In isolated rat liver mitochondria the apparent Km is 52 ยตM, corresponding to 50 % of OXPHOS capacity. The required ADP concentration needed to evaluate Jmax should be minimum 9 times the Km (Gnaiger 2001 Respir Physiol).