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A Combined, Affordable Solution for the Performance of Automated 3D in vitro Hepatotoxicity TestingDownload
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February 13, 2017
Authors: Brad Larson, BioTek Instruments, Inc. Winooski, VT USA; Alexa Rosenberg, Kristina Klette, Michael Mouradian, Hamilton Company, Reno, Nevada USA; Paul Cizdziel, ReproCELL, Inc. Yokohama JP; Glauco R. Souza, Nano3D Biosciences, Inc. Houston, TX USA, The University of Texas Health Science Center at Houston, Houston, TX USA
Prescription medications, environmental toxins, and non-prescription herbal remedies together form the major causes of hepatic injury. When looking at drugs alone, induced hepatic injury is the most common reason cited for warnings or withdrawal of an approved drug, depending on the severity of the induced hepatotoxicity. Due to this reality, a paradigm shift has taken place in the way toxicology studies are being performed, including determination of hepatotoxicity. Early phase assessment of potential hepatotoxic effects from a wider range of new and existing drugs or chemicals is now the norm. Coupled with the change in the development phase where testing takes place, is the desire to use cell models that close the gap between in vitro and in vivo systems.
Hepatotoxicity studies historically were performed by repeatedly dosing hepatocytes cultured on the bottom of a microplate with multiple concentrations of a test drug or compound. Because hepatocytes rapidly dedifferentiate, lose metabolic activity, and lack the communication networks found in vivo when cultured in this manner, results may be inaccurate and yield misleading claims regarding the safety of the test agent. To combat this shortfall, three-dimensional (3D) spheroidal models, such as those using hiPSC-hepatocytes, can be incorporated that allow cells to aggregate and retain typical long-term viability, functionality and communication found in vivo; allowing for the generation of repeatable, accurate data.
To meet the demand for increased hepatotoxicity testing, automation has been incorporated to streamline the procedure and reduce the need for large scale manual manipulations. Typically, included liquid handling systems have been large, expensive, and many times required placement into clean rooms for sterile processing. While this type of solution is suitable for pharmaceutical, biotech, and even larger core facilities, the size and cost can be prohibitive to the typical academic research lab. Therefore, a smaller, less expensive instrumentation set, which can still provide accurate and repeatable results, is necessary.
Here we demonstrate the ability to combine liquid handling with a novel cell imaging multi-mode reader to perform 3D hepatotoxicity studies. hiPSC-hepatocytes were aggregated into spheroids easily and efficiently using magnetic bioprinting technology. Multiple known hepatotoxicants were tested with the automation and cell model to validate the ability of the combined solution to be used to perform automated, walk-away hepatotoxicity studies in an academic setting.