Viability, attachment efficiency, and xenobiotic metabolizing enzyme activities are well maintained in EDTA isolated rat liver parenchymal cells after hypothermic preservation for up to 3 days in University of Wisconsin solution.
Rat liver parenchymal cells were isolated by EDTA perfusion and were subsequently purified by Percoll centrifugation. The freshly isolated liver cells had a mean viability of 95% as judged by trypan blue exclusion. Isolated liver parenchymal cells were then stored at 0 degrees C for up to 1 wk in University of Wisconsin solution (UW). During this hypothermic preservation, the viability was only slightly reduced to 92% after 1 d and to 85% after 3 d at 0 degrees C. Thereafter, the viability decreased rapidly. After cold storage for up to 3 d, it was possible to use the parenchymal liver cells either in short-term suspension or in cell culture. The attachment efficiency in cell culture was the same for freshly isolated liver cells (84%) and after 2 d cold preservation (81%). The cytochrome P450 content and the enzyme activities of soluble epoxide hydrolase, UDP-glucuronosyl transferase, phenol sulfotransferase, and glutathione S-transferase were not significantly different between freshly isolated cells and cells after 3 d of hypothermic preservation. Furthermore, freshly isolated and intact liver cells stored for 3 d were used in the cell-mediated Salmonella mutagenicity test as a metabolizing system. Both fresh and stored liver parenchymal cells metabolized benzo(a)pyrene,2-aminoanthracene, and cyclophosphamide to their ultimate mutagens.(ABSTRACT TRUNCATED AT 250 WORDS)
Institute of Toxicology, University of Mainz, Germany.
F
F
Oesch
H
H
Abdel-Latif
B
B
Diener
D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002448:Cell Adhesion; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004492:Edetic Acid / Q000494:pharmacology; D004798:Enzymes / Q000378:metabolism; D005978:Glutathione / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D008297:Male; D019188:Organ Preservation Solutions; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015262:Xenobiotics / Q000378:metabolism
D003451:Cryoprotective Agents; D004798:Enzymes; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D015262:Xenobiotics; D000493:Allopurinol; D004492:Edetic Acid; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose