Tissue storage
Cold storage
Small tissue samples (10 mg wet weight) should be put immediately into a pre-cooled storage medium, e.g. BIOPS, and stored on ice (0 to 4 °C). Isolated mitochondria should be stored at high concentration (50 mg mt-protein/ml) on ice in mitochondrial preservation medium.[1]
Cold storage time
Up to 12 hours of cold storage does normally not affect mitochondrial respiratory function of liver (pig: 24 h)[2] and human muscle biopsies (skeletal muscle: 24 h[3]; cardiac muscle: 8-12 h[4]).
Muscle biopsies of horses can be stored for 7 days without loss of function (MiPNet12.23 FibreRespiration).
Storage time without loss of function is, therefore, tissue and species dependent and should be evaluated experimentally. When a larger tissue sample is available, separate the sample into small (10 mg) pieces, and apply respirometric SUIT protocols on subsamples in a time course. In particular, evaluate dyscoupling (L/E and P/E coupling control ratios), cytochrome c release,[5] and OXPHOS capacities with various substrate combinations.
References
- ↑ Gnaiger E, Kuznetsov AV, Schneeberger S, Seiler R, Brandacher G, Steurer W, Margreiter R (2000) Mitochondria in the cold. In: Life in the Cold (Heldmaier G, Klingenspor M, eds) Springer, Heidelberg, Berlin, New York: pp 431-442.
- ↑ Kuznetsov AV, Strobl D, Ruttmann E, Königsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Analyt. Biochem. 305: 186-194.
- ↑ Skladal D, Sperl W, Schranzhofer R, Krismer M, Gnaiger E, Margreiter R, Gellerich FN (1994) Preservation of mitochondrial functions in human skeletal muscle during storage in high energy preservation solution (HEPS). In: What is Controlling Life? (Gnaiger E, Gellerich FN, Wyss M, eds) Modern Trends in BioThermoKinetics 3. Innsbruck Univ. Press: 268-271.
- ↑ Lemieux H, Semsroth S, Antretter H, Hoefer D, Gnaiger E (2011) Mitochondrial respiratory control and early defects of oxidative phosphorylation in the failing human heart. Int. J. Biochem. Cell Biol. 43: 1729–1738.
- ↑ Kuznetsov AV, Schneeberger S, Seiler R, Brandacher G, Mark W, Steurer W, Saks V, Usson Y, Margreiter R, Gnaiger E (2004) Mitochondrial defects and heterogeneous cytochrome c release after cardiac cold ischemia and reperfusion. Am. J. Physiol. Heart Circ. Physiol. 286: H1633–H1641.
