title: Reactive oxygen species and cytotoxicity in rainbow trout hepatocytes: effects of medium and incubation time.
abstract: This study evaluated the effects of exposure medium and culture age on intracellular reactive oxygen species (ROS) development and cytotoxicity in fish hepatocytes following exposure to copper (Cu). ROS was quantified using the fluorescent probes DHR 123 and CM-H2DCFDA following exposure to Cu in Leibovitz' medium (L-15) or Tris-buffered saline (TBS). Similarly, culture age effects were investigated using 1-, 2- and 4-day-old cultured hepatocytes by exposing them to Cu in TBS. The exposure in L-15 resulted in significantly higher ROS compared to TBS using CM-H2DCFDA, but not DHR 123. The age of the primary cultures significantly affected the development of ROS for both probes. None of the exposures caused cytotoxicity in the hepatocytes. The results showed that both factors may affect responses to stressors, and suggested that the use of a simple medium such as TBS may be preferable for some applications. It is also preferable to use 1-day-old primary hepatocyte cultures.
authors.affiliation: Department of Biosciences, University of Oslo, Blindern, P.O. Box 1066, N-0316, Oslo, Norway, mazyar.yazdani.edu@gmail.com.
authors.forename: Mazyar
authors.initials: M
authors.lastname: Yazdani
authors.forename: Ragnhild Elisabeth
authors.initials: RE
authors.lastname: Paulsen
authors.forename: Tor
authors.initials: T
authors.lastname: Gjøen
authors.forename: Ketil
authors.initials: K
authors.lastname: Hylland
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003300:Copper / Q000633:toxicity*; D003470:Culture Media; D004785:Environmental Pollutants / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D017686:Oncorhynchus mykiss; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D003470:Culture Media; D004785:Environmental Pollutants; D017382:Reactive Oxygen Species; D003300:Copper