PhysioMimix – Disease Models

  • 3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B virus infection

    With more than 240 million people infected, hepatitis B virus (HBV) is a major health concern. The inability to mimic the complexity of the liver using cell lines and regular primary human hepatocyte (PHH) cultures pose significant limitations for studying host/pathogen interactions. Here, we describe a 3D microfluidic PHH system permissive to HBV infection, which can be maintained for at least 40 days.

    This system enables the recapitulation of all steps of the HBV life cycle, including the replication of patient-derived HBV and the maintenance of HBV cccDNA. We show that innate immune and cytokine responses following infection with HBV mimic those observed in HBV-infected patients, thus allowing the dissection of pathways important for immune evasion and validation of biomarkers. Additionally, we demonstrate that the co-culture of PHH with other non-parenchymal cells enables the identification of the cellular origin of immune effectors, thus providing a valuable preclinical platform for HBV research.

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  • Three-dimensional perfused human in vitro model of nonalcoholic fatty liver disease

    Aim to develop a human in vitro model of non-alcoholic fatty liver disease (NAFLD), utilising primary hepatocytes cultured in a three-dimensional (3D) perfused platform.

    Fat and lean culture media were developed to directly investigate the effects of fat loading on primary hepatocytes
    cultured in a 3D perfused culture system. Oil Red O staining was used to measure fat loading in the hepatocytes and the consumption of free fatty acids (FFA) from culture medium was monitored. Hepatic functions, gene expression profiles and adipokine release were compared for cells cultured in fat and lean conditions. To determine if fat loading in the system could be modulated hepatocytes were treated with known anti-steatotic compounds.

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  • Spontaneous dormancy of metastatic breast cancer cells in an all human liver microphysiologic system.

    Background:

    Metastatic outgrowth in breast cancer can occur years after a seeming cure. Existing model systems of dormancy are
    limited as they do not recapitulate human metastatic dormancy without exogenous manipulations and are unable to query early events of micrometastases.

    Methods:

    Here, we describe a human ex vivo hepatic microphysiologic system. The system is established with fresh human hepatocytes and non-parenchymal cells (NPCs) creating a microenvironment into which breast cancer cells (MCF7 and MDA-MB-231) are added.

    Results:

    The hepatic tissue maintains function through 15 days as verified by liver-specific protein production and drug
    metabolism assays. The NPCs form an integral part of the hepatic niche, demonstrated within the system through their
    participation in differential signalling cascades and cancer cell outcomes. Breast cancer cells intercalate into the hepatic niche without interfering with hepatocyte function. Examination of cancer cells demonstrated that a significant subset enter a quiescent state of dormancy as shown by lack of cell cycling (EdU or Ki67). The presence of NPCs altered the cancer cell fraction entering quiescence, and lead to differential cytokine profiles in the microenvironment effluent.

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