To measure cytochrome c oxidase activity in tissues, the brain and spleen were obtained from sham-operated rats and surviving rats at 72 h post-CA in the vehicle, frozen-thawed, or fresh-mito groups. The COX activity in tissue homogenates was measured using the Cytochrome C oxidase Kit according to the manufacturer’s instructions.
In a separate set of experiments, rats subjected to CA were used to ascertain the uptake and persistence after 1 and 24 h of labeled mitochondrial in vital organs using a confocal microscope. The freshly isolated mitochondria were labeled with MitoTracker Deep Red immediately after isolation, and the vehicle or the labeled mitochondria were infused upon resuscitation from CA.
After tissue isolation, the exogenous donor mitochondria were stained with MitoTracker Deep Red and then co-cultured with neural cells, whose endogenous mitochondria are stained with MitoTracker Green. Exogenous brain mitochondria were taken up into neural cells by simple co-culture for 24 h (Fig.
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