期刊
JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY
卷 27, 期 3, 页码 204-212出版社
WILEY-BLACKWELL
DOI: 10.1002/jbt.21469
关键词
Cocultures; Micropatterning; Rat Hepatocytes; Sandwich Cultures; Stromal Cells
资金
- Long-Range Research Initiative of the American Chemistry Council (ACC)
Primary hepatocytes display functional and structural instability in standard monoculture systems. We have previously developed a model in which primary hepatocytes are organized in domains of empirically optimized dimensions and surrounded by murine embryonic fibroblasts (HepatoPac). Here, we assess the long-term phenotype of freshly isolated and cryopreserved rat hepatocytes in a 96-well HepatoPac format. The viability, cell polarity (actin microfilaments, bile canaliculi), and functions (albumin, urea, Phase I/II enzymes, transporters) of fresh and cryopreserved rat hepatocytes were retained in HepatoPac at similar levels for at least 4 weeks as opposed to rapidly declining over 5 days in collagen/Matrigel sandwich cultures. Pulse or continuous exposure of rat HepatoPac to GW-7647, a selective agonist of PPAR, caused reproducible induction of CYP4A1 and 3-hydroxy-3-methylglutaryl-CoA synthase over 4 weeks. In conclusion, rat HepatoPac in a 96-well format can be used for chronic dosing of highly functional hepatocytes and assessment of perturbed hepatocellular pathways. (c) 2013 Wiley Periodicals, Inc. J BiochemMol Toxicol 27:204-212, 2013; View this article online at wileyonlinelibrary.com. DOI 10.1002/jbt.21469
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