Tissue storage: Difference between revisions
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==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 Β°C). | |||
==Cold storage time== | |||
Up to 12 hours of cold storage does normally not affect mitochondrial respiratory function of liver (pig: 24 h)<ref name ="Kuznetsov 2002 AnalytBiochem">[[Kuznetsov 2002 AnalytBiochem|Kuznetsov AV, Strobl D, Ruttmann E, KΓΆnigsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Analyt. Biochem. 305: 186-194]].</ref> and human muscle biopsies (skeletal muscle: 24 h<ref name ="Skladal 1994 BTK">[[Skladal 1994 BTK|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]].</ref>; cardiac muscle: 8-12 h<ref name ="Lemieux 2011 IJBCB">[[Lemieux 2011 IJBCB|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: 1729β1738]].</ref>). Β | |||
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 protocol]]s on subsamples in a time course. In particular, evaluate [[dyscoupling]] (''L/E'' and ''P/E'' coupling control ratios), cytochrome c release,<ref name ="Kuznetsov 2004 AJP">[[Kuznetsov 2004 AJP|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: H1633βH1641]].</ref> and [[OXPHOS]] capacities with various substrate combinations. | |||
==References== | |||
<references/> | |||
Revision as of 18:51, 12 November 2011
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 Β°C).
Cold storage time
Up to 12 hours of cold storage does normally not affect mitochondrial respiratory function of liver (pig: 24 h)[1] and human muscle biopsies (skeletal muscle: 24 h[2]; cardiac muscle: 8-12 h[3]).
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,[4] and OXPHOS capacities with various substrate combinations.
References
- β Kuznetsov AV, Strobl D, Ruttmann E, KΓΆnigsrainer A, Margreiter R, Gnaiger E (2002) Evaluation of mitochondrial respiratory function in small biopsies of liver. Analyt. Biochem. 305: 186-194.
- β 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.
- β 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: 1729β1738.
- β 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: H1633βH1641.