title: Isolation of mitochondria from tissue culture cells.
abstract: The number of mitochondria per cell varies substantially from cell line to cell line. For example, human HeLa cells contain at least twice as many mitochondria as smaller mouse L cells. This protocol starts with a washed cell pellet of 1-2 mL derived from ∼10⁹ cells grown in culture. The cells are swollen in a hypotonic buffer and ruptured with a Dounce or Potter-Elvehjem homogenizer using a tight-fitting pestle, and mitochondria are isolated by differential centrifugation.
authors.affiliation: Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147-2408;
authors.forename: David A
authors.initials: DA
authors.lastname: Clayton
authors.affiliation: Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520-8023; Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8023.
authors.forename: Gerald S
authors.initials: GS
authors.lastname: Shadel
mesh_terms: D000818:Animals; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured / Q000648:ultrastructure*; D002498:Centrifugation; D006801:Humans; D051379:Mice; D008928:Mitochondria / Q000502:physiology*