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Comparison of hepatocytes in monolayer and RAFT™ 3D Cell Culture System
EP29425
Poster Title: Comparison of hepatocytes in monolayer and RAFT™ 3D Cell Culture System
Submitted on 29 Nov 2018
Author(s): Therese Willstaedt, Maureen Bunger, Lubna Hussain, and Theresa D’Souza
Affiliations: Lonza
This poster was presented at SOT 2018
Poster Views: 515
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Poster Information
Abstract: Hepatocytes comprise 70 – 85% of the liver’s mass and are the major cell type involved in detoxification. In the pharmaceutical industry there is an increasing demand for hepatocyte-based models which mimic human in vivo toxicity. For in-vitro screening, the current gold standard for drug metabolism and toxicity consists of hepatocytes grown is a traditional two dimensional (2D) culture system. However, it has been reported that although primary hepatocytes briefly maintain some functional capabilities in monolayer culture, such as the production of drug-metabolizing enzymes, they rapidly lose their morphology along with the majority of liver-specific functions. We compared cell viability, cell morphology and ZO-1 expression of primary rat and human hepatocytes grown as a monolayer on collagen and sandwiched by Matrigel with that of cells cultured within collagen hydrogel in three dimensions using RAFT™ Culture System. To determine the effect of growing hepatocytes in RAFT™ Culture compared to sandwich culture, we treated the human hepatocytes in both models with inducers like Omeprazole, Phenobarbitol and Rifampycin and examined the metabolic cytochrome P450 (CYP) capabilities. For hepatocytes in RAFT™ Culture System the response was both stronger and more stable over 17 days in culture compared to the sandwich cultures. Improved stability of P450s by culturing hepatocytes in 3 Dimensions could enable long-term and repeat dosing for toxicity analysis using primary human hepatocytes.

Summary: In this poster we compare cell viability and cell morphology of rat and human hepatocytes, and the maintenance of Cytochrome P450 (CYP) activity in human hepatocytes grown in the traditional Sandwich Model with that of cells cultured in the 3D RAFT™ System. Our results show that the RAFT™ 3D System represents a more robust model for the long-term maintenance of liver-specific functions.Report abuse »
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