caa a22 4500 477081614 CHVBK 20180405111451.0 cr unu---uuuuu 170330e19960401xx s 000 0 eng 10.1007/BF02722948 doi (NATIONALLICENCE)springer-10.1007/BF02722948 Characterization of a primary hepatocyte culture system for toxicological studies [Elektronische Daten] [Joanne Zurlo, Linda Arterburn] Summary: An hepatocyte culture system was developed for potential use in toxicological studiesin vitro. Rat hepatocytes were isolated by two-step collagenase perfusion and cultured on Vitrogen-coated Permanox™ dishes in a modified Chee's medium containing 1μM dexamethasone and 1% dimethylsulfoxide. The cells remained highly viable for at least 10 d as determined by lactate dehydrogenase release and total protein levels. Albumin secretion into the medium, as a measure of differentiated function, was maintained at elevated levels over the course of 10 d in culture. A number of CYP activities were determined by the analysis of testosterone metabolism in freeze-thawed cells, diazepam metabolism in live cells, and specific assays for CYP 1A1/2, 2B1/2, 2E1, and 3A. Results of these assays indicated that a wide range of CYP isozymes were maintained, some activities were enhanced under the conditions of culture and some activities were inducible. Activities of the phase II enzymes, glutathione S-transferase and UDP-glucuronosyltransferase, and glutathione levels were also maintained in the cultured hepatocytes for at least 6 d. These results strongly support the use of this hepatocyte culture system forin vitro toxicological studies. Society for In Vitro Biology, 1996 rat hepatocytes nationallicence in vitro toxicology nationallicence metabolism nationallicence cytochrome P450 nationallicence Zurlo Joanne Division of Toxicological Sciences and In Vitro Toxicology Program of the Johns Hopkins Center for Alternatives to Animal Testing, 111 Market Place, Suite 840, 21202-6709, Baltimore, Maryland aut Arterburn Linda Research Division, W. R. Grace & Co.-Conn., Columbia, Maryland aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/4(1996-04-01), 211-220 1071-2690 32:4<211 1996 32 11626 https://doi.org/10.1007/BF02722948 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02722948 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zurlo Joanne Division of Toxicological Sciences and In Vitro Toxicology Program of the Johns Hopkins Center for Alternatives to Animal Testing, 111 Market Place, Suite 840, 21202-6709, Baltimore, Maryland aut NATIONALLICENCE 700 1- Arterburn Linda Research Division, W. R. Grace & Co.-Conn., Columbia, Maryland aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 32/4(1996-04-01), 211-220 1071-2690 32:4<211 1996 32 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer