Isolation of Intact Mitochondria from Skeletal Muscle by Differential Centrifugation for High-resolution Respirometry Measurements.

Djafarzadeh, Siamak; Jakob, Stephan (2017). Isolation of Intact Mitochondria from Skeletal Muscle by Differential Centrifugation for High-resolution Respirometry Measurements. Journal of visualized experiments(121) MYJoVE Corporation 10.3791/55251

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Mitochondria are involved in cellular energy metabolism and use oxygen to produce energy in the form of adenosine triphosphate (ATP). Differential centrifugation at low- and high-speed is commonly used to isolate mitochondria from tissues and cultured cells. Crude mitochondrial fractions obtained by differential centrifugation are used for respirometry measurements. The differential centrifugation technique is based on the separation of organelles according to their size and sedimentation velocity. The isolation of mitochondria is performed immediately after tissue harvesting. The tissue is immersed in an ice-cold homogenization medium, minced using scissors and homogenized in a glass homogenizer with a loose-fitting pestle. The differential centrifugation technique is efficient, fast and inexpensive and the mitochondria obtained by differential centrifugation are pure enough for respirometry assays. Some of the limitations and disadvantages of isolated mitochondria, based on differential centrifugation, are that the mitochondria can be damaged during the homogenization and isolation procedure and that large amounts of the tissue biopsy or cultured cells are required for the mitochondrial isolation.

Item Type:

Journal Article (Further Contribution)

Division/Institute:

04 Faculty of Medicine > Department of Intensive Care, Emergency Medicine and Anaesthesiology (DINA) > Clinic of Intensive Care
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Pavillon 52 > Forschungsgruppe Intensivmedizin

UniBE Contributor:

Djafarzadeh, Siamak and Jakob, Stephan

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1940-087X

Publisher:

MYJoVE Corporation

Language:

English

Submitter:

Mirella Aeberhard

Date Deposited:

27 Feb 2018 11:00

Last Modified:

27 Feb 2018 11:00

Publisher DOI:

10.3791/55251

PubMed ID:

28362420

URI:

https://boris.unibe.ch/id/eprint/109759

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