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Determination of metabolic stability using cryopreserved hepatocytes from rainbow trout (Oncorhynchus mykiss)
Citation:
Fay, K., D. Nabb, R. Mingoia, I. Bischoff, H. Segner, X. Han, AND K. Johanning. Determination of metabolic stability using cryopreserved hepatocytes from rainbow trout (Oncorhynchus mykiss). Current Protocols in Toxicology. Wiley InterScience, Silver Spring, MD, 65:4.42.1-4.42.29, (2015).
Impact/Purpose:
Increasingly, in vitro systems derived from liver tissue are being used to measure chemical biotransformation rates for fish. This information is then being used to predict metabolism impacts on chemial accumulation. The use of such methods within a regulatory context requires, however, that they be rigorously standardized. This contribution provides detailed methods for isolating and cryopreserving hepatocytes from rainbow trout. Additional instructions for use of this system to measure metabolic activity in fish and extrapolate this information to the intact animal are provided. This paper provides critical and timely guidance on the use of an important in vitro system for bioaccumulation assessments with fish.
Description:
Standard protocols for isolating, cryopreserving, and thawing rainbow trout hepatocytes are described, along with procedures for using fresh or cryopreserved hepatocytes to assess chemical metabolic stability 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 species with appropriate considerations.
