RAPID METHOD FOR THE ISOLATION OF INTACT MITOCHONDRIA FROM ISOLATED RAT-LIVER CELLS

A method is described for the preparation of intact mitochondria from isolated hepatocytesby sonication. Sonication of a suspension of rat liver cells for 10-30 s yields a homogenate from which tightly coupled mitochondria can be isolated. These mitochondria exhibit high respiratory control ratios and normal ADP:O ratios using glutamate plus malate, β-hydroxybutyrate, succinate, or ascorbate plus N,N,N′,N′-tetramethyl-p-phenylendiamine as substrates. The yield of mitochondrial protein is approximately 100-120 mg starting from 5 g of liver tissue. The mitochondrial fraction is essentially free of contaminating plasma membrane and microsomes and contains only small amounts of peroxisomes and lysosomes. © 1979 Academic Press, Inc.