This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Grant, M I Articles by MacDonald, C Search for Related Content PubMed PubMed Citation Articles by Grant, M I Articles by MacDonald, C Social Bookmarking                   What's this?

Manipulation of the phenotype of immortalised rat hepatocytes by different culture configurations and by dimethyl sulphoxide

K Anderson

G McKay

M Wills

C Henderson

Bioengineering Unit, Strathclyde University, Wolfson Centre, 106 Rottenrow, Glasgow G4 ONW, UK

C MacDonald

Biological Sciences, University of Paisley, High Street, Paisley PAl 2BE, UK

The liver-specific phenotype of immortalised rat hepato-cytes is not irretrievably lost as they age in culture but can be manipulated by modifying the culture environment.

Testosterone metabolism was used to investigate the profile of cytochrome P450 isoenzymes present in two immortalised cell lines, P9 and LQC, and in primary cultures of rat hepatocytes, cultured on collagen films, gels and double gel cultures (sandwich configuration). The extent of testosterone metabolism, and the range of metabolites produced, was increased in immortalised cells by the presence of collagen as a substratum film or gel but survival was poorer and the range of metabolites was reduced in sandwich culture. In contrast, testosterone metabolism was retained in primary hepatocytes in sandwich cultures at a higher level than in collagen film or gel cultures.

Expression of alpha class glutathione-S-transferases (GSTs) increased and that of GSTP1 decreased (changes which indicate a recovery of normal liver GST phenotype) when the medium of immortalised cell cultures was supplemented with dimethyl sulph-oxide (DMSO). DMSO also improved ethoxyresorufin 0-deethylation (EROD) and testosterone metabolism in immortalised cells. It also markedly inhibited prolifera-tion, DNA, RNA and protein synthesis.

Maximal testosterone metabolism was observed in immortalised cells cultured on collagen gels in the presence of l1o (v/v) DMSO. Development of a protocol for treating immortalised liver cells cultured on collagen gels with DMSO to switch between proliferation and differentiation may provide a convenient system expres-sing the xenobiotic metabolising enzymes required for in vitro toxicity testing.

Key Words: immortalised hepatocytes • collagen substrata • culture configuration • dimethyl sulphoxide

Human & Experimental Toxicology, Vol. 19, No. 5, 309-317 (2000)
DOI: 10.1191/096032700678815936


CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Kataropoulou, C. Henderson, and H. Grant The influence of glycosaminoglycans and crosslinking agents on the phenotype of hepatocytes cultured on collagen gels Human and Experimental Toxicology, February 1, 2003; 22(2): 65 - 71. [Abstract] [PDF]