Isolation of Mitochondria from Tissue Culture Cells | Zendy

David A. Clayton | Zendy; Gerald S. Shadel | Zendy

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Isolation of Mitochondria from Tissue Culture Cells

Author(s) -

David A. Clayton,

Gerald S. Shadel

Publication year - 2014

Publication title -

cold spring harbor protocols

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.674

H-Index - 51

eISSN - 1940-3402

pISSN - 1559-6095

DOI - 10.1101/pdb.prot080002

Subject(s) - hela , mitochondrion , cell culture , homogenizer , differential centrifugation , centrifugation , microbiology and biotechnology , tonicity , cell , isolation (microbiology) , chemistry , tissue culture , biology , biochemistry , genetics , in vitro

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.

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