Isolation of Mitochondria from Tissue Culture Cells
- 1Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147-2408;
- 2Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520-8023;
- 3Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8023
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.
Footnotes
-
↵4 Correspondence: gerald.shadel{at}yale.edu
- © 2014 Cold Spring Harbor Laboratory Press
