Abstract
The processes of modern drug discovery and development rely on the ability to obtain information regarding new chemical entities as quickly and inexpensively as possible. For this reason, laboratories have developed various in vitro techniques that can help to minimize undue investment in developmental compounds that may have undesirable pharmacokinetic properties and/or the potential to cause adverse effects, such as toxicity and drug–drug interactions. The primary culture of human hepatocytes offers a reliable in vitro system to test a compound’s ability to induce the expression of cytochrome P450 enzymes, a primary route of metabolism for many pharmaceuticals. The methods in this chapter describe the isolation of primary hepatocytes from nontransplantable human liver followed by their culture and treatment with new chemical entities and/or known inducers of cytochrome P450. Enzyme induction in cultured human hepatocytes can be assessed by measuring the levels of messenger ribonucleic acid, immunoreactive protein, and/or cytochrome P450 enzyme activity as outlined in this chapter.
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