Project: Liver spheroids: An advanced HTS in vitro system for the study of drug induced liver toxicity and liver diseases.
InSphero has developed a 3D liver model produced from cryopreserved human hepatocytes in co-culture with non-parenchymal cells, including endothelial and Kupffer cells.The so-called liver microtissues are produced by InSphero’s patented protected hanging drop culture format, designed to obtain regularly shaped scaffold-free liver spheroids provided in a 96-well standard microtiter plate format amenable to standard liquid handling robotic systems. The spheroids are highly uniform in shape and size and show a morphology which is close to the native liver tissue, with non-parenchymal cells, COly endothelial cells and Kupffer cells interspersed between hepatocytes. When cultured in InSphero’s GravityTRAP format, which was specifically designed for long term spheroid culture, microtissue viability and liver specific properties, such as urea production, albumin secretion and CYP3A activity can be sustained for at least 4 weeks._x000D__x000D_This advanced 3D liver model has already been demonstrated to be superior to conventional 2D hepatocyte cultures, not only with respect to long term viability and functionality but also by its ability to mimic inflammation mediated toxicity through the activation of Kupffer cells. The systems is compatible to the same endpoint measurements commonly used in liver toxicology, such as biochemical assays (viability/cell death/apoptosis/oxidative stress), metabolomics, enzyme activity, and high content analysis._x000D__x000D_As the proposed in vitro liver model system has been characterized to fulfill fundamental requirements with respect to the ability to be implemented into current toxicity screening routines ( i.e. adhering to standard cell culture formats, its compatibility with liquid handling systems and the robustness of data acquired), the project aims to validate the model in terms of its predictive power in comparison to state of the art in vitro 2D liver toxicity models. This will include whole genome gene expression analysis, phase I and II metabolizing enzyme activity profiling. Common drug induced liver toxicities will be simulated in order to confirm the corresponding biochemical and morphological hallmarks and the underlying mechanisms. Other endpoints relevant to drug induced acute and chronic liver toxicity, such as hypertrophy, cholestasis or fibrosis will be established and validated. In order to establish the predictive power of this model system, toxicity will be assessed in the liver microtissues exposed to a panel of selected known hepatotoxic compounds, including those withdrawn from the market. _x000D_ _x000D__x000D_In parallel, the development of a 384-well hanging drop plate format based on InSphero's core technology shall assure high-throughput capabilities and a streamlined processing of organotypic microtissues, from production to long term culture and treatment procedures. As this new in vitro liver toxicity model also offers analysis by classical histological methods, the projects aims to develop a device for simultaneous embedding of multiple tisssue specimens which will allow higher throughput capacities in microtissue processing for histological assessments._x000D__x000D_The project will serve two major aims:_x000D__x000D_1) To generate a a comprehensive data set as a compelling marketing concept in order to persuade customers in the pharmaceutical, cosmetic and chemical industry to adapt to the new in vitro liver toxicity model for a more reliable and predictive safety assessment of drugs during the pre-clinical phase of drug development. _x000D__x000D_2) To advance InSphero's current technology to an even higher throughput system, which will offer a 384-well format for the production and long-term culture of liver microtissues and downstream processing assays, including histology._x000D_
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Acronym
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HTS DILI
(Reference Number: 8511)
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Duration
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01/11/2013 - 31/01/2016
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Project Topic
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This project involves an in-depth validation of a newly developped scaffold-free 3D human liver culture model for the detectionof drug induced liver toxicity and its upscaling to a 384-well format. We will develop chronic tox testing strategies and also establish rare liver tox specific endpoints.
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Network
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Eurostars
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Call
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Eurostars Cut-Off 10
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Project partner
