title: Determination of Metabolic Stability Using Cryopreserved Hepatocytes from Rainbow Trout (Oncorhynchus mykiss).
abstract: Trout provide a relatively easy source of hepatocytes that can be cryopreserved and used for a range of applications including toxicity testing and determination of intrinsic clearance. Standard protocols for isolating, cryopreserving, and thawing rainbow trout hepatocytes are described, along with procedures for using fresh or cryopreserved hepatocytes to assess metabolic stability of xenobiotics in fish by means of a substrate depletion approach. Variations on these methods, troubleshooting tips, and directions for use of extrapolation factors to express results in terms of in vivo intrinsic clearance are included. These protocols have been developed for rainbow trout, but can be adapted to other fish species with appropriate considerations.
authors.affiliation: ORD/NHEERL/Mid-Continent Ecology Division, U.S. EPA, Duluth, Minnesota.
authors.forename: Kellie A
authors.initials: KA
authors.lastname: Fay
authors.affiliation: DuPont Haskell Global Centers for Health and Environmental Sciences, Newark, Delaware.
authors.forename: Diane L
authors.initials: DL
authors.lastname: Nabb
authors.affiliation: DuPont Haskell Global Centers for Health and Environmental Sciences, Newark, Delaware.
authors.forename: Robert T
authors.initials: RT
authors.lastname: Mingoia
authors.affiliation: Centre for Fish and Wildlife Health, University of Bern, Bern, Switzerland.|Fraunhofer Institute for Molecular Biology and Applied Ecology, Schmallenberg, Germany.
authors.forename: Ina
authors.initials: I
authors.lastname: Bischof
authors.affiliation: ORD/NHEERL/Mid-Continent Ecology Division, U.S. EPA, Duluth, Minnesota.
authors.forename: John W
authors.initials: JW
authors.lastname: Nichols
authors.affiliation: Centre for Fish and Wildlife Health, University of Bern, Bern, Switzerland.
authors.forename: Helmut
authors.initials: H
authors.lastname: Segner
authors.affiliation: KJ Scientific LCC, Texas Life Sciences Collaboration Center, Georgetown, Texas.
authors.forename: Karla
authors.initials: K
authors.lastname: Johanning
authors.affiliation: DuPont Haskell Global Centers for Health and Environmental Sciences, Newark, Delaware.
authors.forename: Xing
authors.initials: X
authors.lastname: Han
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001711:Biotransformation / Q000502:physiology; D002478:Cells, Cultured; D015925:Cryopreservation / Q000662:veterinary*; D004734:Energy Metabolism / Q000502:physiology*; D022781:Hepatocytes / Q000378:metabolism*; D017686:Oncorhynchus mykiss
keywords: biotransformation; fish; hepatocytes; in vitro assay; metabolic stability; rainbow trout; substrate depletion