西亚试剂：Oxygen-mediated enhancement of primary hepatocyte metabolis

发布时间：2025-10-09

Oxygen-mediated enhancement of primary hepatocyte metabolism, functional polarization, gene expression, and drug clearance

Srivatsan Kidambia, Rubin S. Yarmusha, Eric Novikb, Piyun Chaob, Martin L. Yarmusha and Yaakov Nahmiasa,c,1

aCenter for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02111;
bHμREL Corporation, Beverly Hills, CA 90211; and
cThe Selim and Rachel Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem 91904, Israel

The liver is a major site for the metabolism of xenobiotic compounds due to its abundant level of phase I/II metabolic enzymes. With the cost of drug development escalating to over $400 million/drug there is an urgent need for the development of rigorous models of hepatic metabolism for preclinical screening of drug clearance and hepatotoxicity. Here, we present a microenvironment in which primary human and rat hepatocytes maintain a high level of metabolic competence without a long adaptation period. We demonstrate that co-cultures of hepatocytes and endothelial cells in serum-free media seeded under 95% oxygen maintain functional apical and basal polarity, high levels of cytochrome P450 activity, and gene expression profiles on par with freshly isolated hepatocytes. These oxygenated co-cultures demonstrate a remarkable ability to predict in vivo drug clearance rates of both rapid and slow clearing drugs with an R2 of 0.92. Moreover, as the metabolic function of oxygenated co-cultures stabilizes overnight, preclinical testing can be carried out days or even weeks before other culture methods, significantly reducing associated labor and cost. These results are readily extendable to other culture configurations including three-dimensional culture, bioreactor studies, as well as microfabricated co-cultures.

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