Isolation and culture of hepatocytes from human liver of immediate autopsy

Abstract

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 intack lobe with collagenase or by digestion of a small tissue wedge with collagenase-dispase. The yields of hepatocytes ranged from 1 to 3 × 106 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 cytochromep 450 and 536. pmol/mg protein cytochromeb 5 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.

DOI: 10.1007/BF02621352

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@article{Hsu1985IsolationAC, title={Isolation and culture of hepatocytes from human liver of immediate autopsy}, author={Iawen Hsu and Michael M. Lipsky and Katherine E. Cole and Chun H Su and Benjamin F . Trump}, journal={In Vitro Cellular & Developmental Biology}, year={1985}, volume={21}, pages={154-160} }