Isolation and culture of hepatocytes from human liver of immediate autopsy.
Human livers were removed at immediate autopsy (IA) from brain death patients within 1 h after cessation of cardiac function. Viable hepatocytes were isolated successfully from these IA livers by perfusion of an intact lobe with collagenase or by digestion of a small tissue wedge with collagenase-dispase. The yields of hepatocytes ranged from 1 to 3 X 10(6) cells/g liver in the five cases studied. Approximately 70 to 90% of the cells excluded trypan blue dye. In the isolated hepatocytes, 632 pmol/mg protein of cytochrome p450 and 536 pmol/mg protein cytochrome b5 were measured. The cells attached to the dishes in 4 h and produced monolayer cultures with a high success rate. The cells maintained in primary cultures for several days and developed ultrastructural features characteristic of human hepatocytes in vivo. The cultured hepatocytes can hydroxylate benzo[a]pyrene, conjugate the metabolites, and have a benzo[a]pyrene hydroxylase activity of 48.7 pmol/mg DNA per h, which is comparable to that of rat hepatocytes. The liver cells repaired DNA damage caused by exposures to aminofluorene and acetylaminofluorene in culture.
D000818:Animals; D001344:Autopsy; D001580:Benzopyrenes / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D003573:Cytochrome b Group / Q000032:analysis; D015786:Cytochromes b5; D004247:DNA / Q000032:analysis; D004847:Epithelial Cells; D004848:Epithelium / Q000032:analysis; D006801:Humans; D008099:Liver / Q000166:cytology*; D008297:Male; D008862:Microsomes, Liver / Q000032:analysis; D051381:Rats; D011916:Rats, Inbred F344
D001580:Benzopyrenes; D003573:Cytochrome b Group; C021338:3-hydroxybenzo(a)pyrene; D004247:DNA; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System