caa a22 4500 397556624 CHVBK 20180308164804.0 cr unu---uuuuu 161202e199609 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021443 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021443 Recovery of mRNA Expression of Tryptophan 2,3-Dioxygenase and Serine Dehydratase in Long-Term Cultures of Primary Rat Hepatocytes [Elektronische Daten] [Toru Mizuguchi, Toshihiro Mitaka, Koichi Hirata, Toshikazu Nakamura, Yohichi Mochizuki] Expression of tryptophan 2,3-dioxygenase (TO) and serine dehydratase (SDH) has not previously been maintained or re-induced in long-term cultured hepatocytes. In the present study, we succeeded in inducing expression of TO and SDH mRNAs in adult rat hepatocytes cultured in serum-free L-15 medium supplemented with epidermal growth factor and 2% dimethyl sulfoxide (DMSO). After the start of culture, the expression of TO mRNA rapidly disappeared and at 96 h it was less than 10% of that at isolation. However, after the addition of 2% DMSO from 96 h, the transcript level gradually increased and reached about 40% of that of the isolated cells at day 14. In addition, the expression of TO mRNA was enhanced in cells treated with both 10-5 M dexamethasone and 10-7 M glucagon. In contrast, the expression of SDH mRNA decreased very rapidly and we could not detect it after 24 h of culture. Furthermore, 2% DMSO failed to induce it. However, when both 10-5 M dexameth asone and 10-7 M glucagon were added to the culture medium at day 9, we observed dramatic induction of SDH mRNA 24 h later. Primary hepatocytes cultured by this method could express and maintain highly differentiated hepatic functions for a long time. Thus, this in vitro system is suitable for the investigation of hepatic functions. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence dimethyl sulfoxide nationallicence primary rat hepatocytes nationallicence re-expression nationallicence serine dehydratase nationallicence tryptophan 2,3-dioxygenase nationallicence Mizuguchi Toru Department of Pathology, Cancer Research Institute aut Mitaka Toshihiro Department of Pathology, Cancer Research Institute aut Hirata Koichi Department of Surgery, Section 1, Sapporo Medical University School of Medicine S-1, W-17, Chuo-ku, Sapporo 060 aut Nakamura Toshikazu Biomedical Research Center, Osaka University Medical School aut Mochizuki Yohichi Department of Pathology, Cancer Research Institute aut The Journal of Biochemistry Oxford University Press 120/3(1996-09), 511-517 0021-924X 120:3<511 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021443 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021443 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mizuguchi Toru Department of Pathology, Cancer Research Institute aut NATIONALLICENCE 700 1- Mitaka Toshihiro Department of Pathology, Cancer Research Institute aut NATIONALLICENCE 700 1- Hirata Koichi Department of Surgery, Section 1, Sapporo Medical University School of Medicine S-1, W-17, Chuo-ku, Sapporo 060 aut NATIONALLICENCE 700 1- Nakamura Toshikazu Biomedical Research Center, Osaka University Medical School aut NATIONALLICENCE 700 1- Mochizuki Yohichi Department of Pathology, Cancer Research Institute aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/3(1996-09), 511-517 0021-924X 120:3<511 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford