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Publication Detail

Title: Assessment of compound hepatotoxicity using human plateable cryopreserved hepatocytes in a 1536-well-plate format.

Authors: Moeller, Timothy A; Shukla, Sunita J; Xia, Menghang

Published In Assay Drug Dev Technol, (2012 Feb)

Abstract: Hepatotoxicity is a major concern for both drug development and toxicological evaluation of environmental chemicals. The assessment of compound-induced hepatotoxicity has traditionally relied on in vivo testing; however, it is being replaced by human in vitro models due to an emphasis on the reduction of animal testing and species-specific differences. Since most cell lines and hybridomas lack the full complement of enzymes at physiological levels found in the liver, primary hepatocytes are the gold standard to study liver toxicities in vitro due to the retention of most of their in vivo activities. Here, we optimized a cell viability assay using plateable cryopreserved human hepatocytes in a 1536-well-plate format. The assay was validated by deriving inhibitory concentration at 50% values for 12 known compounds, including tamoxifen, staurosporine, and phenylmercuric acetate, with regard to hepatotoxicity and general cytotoxicity using multiple hepatocyte donors. The assay performed well, and the cytotoxicity of these compounds was confirmed in comparison to HepG2 cells. This is the first study to report the reliability of using plateable cryopreserved human hepatocytes for cytotoxicity studies in a 1536-well-plate format. These results suggest that plateable cryopreserved human hepatocytes can be scaled up for screening a large compound library and may be amenable to other hepatocytic assays such as metabolic or drug safety studies.

PubMed ID: 22053711 Exiting the NIEHS site

MeSH Terms: Cell Culture Techniques/methods*; Cell Survival/drug effects; Cell Survival/physiology; Cells, Cultured; Cryopreservation*/methods; Cytotoxins/toxicity*; Dose-Response Relationship, Drug; Hepatocytes/drug effects*; Hepatocytes/physiology; Humans

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