Hepatocytes from rat liver were prepared by perfusion with collagenase, and rough and smooth microsomes and mitochondria were prepared after cell disruption. By applying 1000 lb/in2 (1 lb/in2 = 6.9 kPa) in a nitrogen bomb followed by decompression, 75% of the cells were disrupted after four consecutive treatments. Intact mitochondria, and rough and smooth microsomes with little contamination were prepared from the homogenate. A more rapid disruption was attained by a short sonication with a low output, thus increasing the efficiency of homogenization. The microsomal subfractions prepared from this homogenate were comparable to those obtained after decompression. Sonication resulted in smooth microsomes, which exhibited a higher contamination with non-microsomal membranes. These, however, were partly removed by additional centrifugation with a discontinuous sucrose gradient containing divalent cations.
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JOURNAL ARTICLE| 01 October 1982
Fractionation of isolated liver cells after disruption with a nitrogen bomb and sonication
Online Issn: 1477-9137
Print Issn: 0021-9533
© 1982 by Company of Biologists
J Cell Sci (1982) 57 (1): 1–13.
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F. Autuori, U. Brunk, E. Peterson, G. Dallner; Fractionation of isolated liver cells after disruption with a nitrogen bomb and sonication. J Cell Sci 1 October 1982; 57 (1): 1–13. doi: https://doi.org/10.1242/jcs.57.1.1
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