Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1007/s11626-998-0030-0
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Titel (primär) Viability and differential function of rainbow trout liver cells in primary culture: Coculture with two permanent fish cells
Autor Scholz, S. ORCID logo ; Braunbeck, T.; Segner, H.
Quelle In Vitro Cellular & Developmental Biology-Animal
Erscheinungsjahr 1998
Department OEC; ZELLTOX; COE
Band/Volume 34
Heft 10
Seite von 762
Seite bis 771
Sprache englisch
Keywords biotransformation; coculture; hepatocytes; fish; continuous cell lines; RTG-2; RTH-149
Abstract The study investigates the influence of different culture conditions on attachment, viability and functional status of rainbow trout (Oncorhynchus mykiss) liver cells in primary culture. Cells were isolated by a two-step collagenase perfusion and incubated in serum-free, chemically defined minimal essential medium (MEM), (a) as a monolayer on uncoated PRI-MARIA® dishes, (b) as a monolayer on culture dishes coated with calf collagen type 1, and (c) in coculture with the established fish cell lines RTH-149 or RTG-2. Cell attachment was assessed from DNA and protein concentrations per dish, viability was estimated from cellular lactate dehydrogenase release, and the metabolic status was investigated by measuring activities of the phosphoenolpyruvate carboxykinase and biotransformation enzymes as well as the total cytochrome P450 contents. Seeding of hepatocytes on collagen-coated dishes did not alter cell attachment or detachment from the culture substrate, but had a small, but not significant effect on cell viability and metabolic parameters. Coculture of liver cells and RTG-2 cells reduced hepatocyte detachment from the culture substrate, and it was associated with a significant elevation of 7-ethoxyresorufin-O-deethylase activities in the hepatic cells. Cytochrome P450 contents, however, were not altered. The coculture effect on liver cell physiology clearly depended on the type of cell line, because coculture with RTH-149 cells led to similar, but much weaker effects than obtained in cocultures with RTG-2 cells. Electron microscopical observations revealed the existence of gap junctions and possible exocytosis-like transport between cell lines and hepatocytes. The results point to the potential of coculture systems to improve physiological parameters of trout liver cells in primary culture.
dauerhafte UFZ-Verlinkung
Scholz, S., Braunbeck, T., Segner, H. (1998):
Viability and differential function of rainbow trout liver cells in primary culture: Coculture with two permanent fish cells
In Vitro Cell. Dev. Biol.-Anim. 34 (10), 762 - 771 10.1007/s11626-998-0030-0