Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts.
Mitochondria participate in key metabolic reactions of the cell and regulate crucial signaling pathways including apoptosis. Although several approaches are available to study mitochondrial function in situ are available, investigating functional mitochondria that have been isolated from different tissues and from cultured cells offers still more unmatched advantages. This protocol illustrates a step-by-step procedure to obtain functional mitochondria with high yield from cells grown in culture, liver and muscle. The isolation procedures described here require 1-2 hours, depending on the source of the organelles. The polarographic analysis can be completed in 1 hour.
Frezza
Christian
C
Dulbecco-Telethon Institute, Venetian Institute of Molecular Medicine, Via Orus 2, 35129 Padova, Italy.
Cipolat
Sara
S
Scorrano
Luca
L
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology*; D008099:Liver / Q000166:cytology*; D051379:Mice; D008928:Mitochondria / Q000648:ultrastructure*; D018482:Muscle, Skeletal / Q000166:cytology*