3D InSight™ Human Liver Microtissues

Delivered ready-to-use in 96-well assay plates 

  • Co-culture of primary human hepatocytes with Kupffer cells
  • Suitable for idiosyncratic and long-term toxicity with repeated dosing
  • Economical and convenient 96-well format (GravityTRAP™)
  • Delivered within 24 hours at 30-37°C
 

  • Organotypic cell composition

    3D InSight™ Human Liver Microtissues are generated by co-culture of primary human hepatocytes and non-parenchymal Kupffer cells to reflect the native cell composition of liver. To assure short delivery times and the availability of specific donor lots over months and years, InSphero has developed a unique production process based on cryopreserved hepatocytes. InSphero's expert-know-how in generation of 3D InSight™ Human Liver Microtissues guarantees excellent functionality, viability and reproducibility.

    3D InSight™ Human Liver Microtissues are delivered as a ready-to-use model in standard 96-well format (GravityTRAP™). This format allows easy cultivation, observation and assaying of 3D InSight™ Human Liver Microtissues, without risk of losing microtissues over the culture time. InSphero's new liver model retains similar multiple endpoint compatibility as the currently used 2D culture formats but significantly enhances the predictive power of such assays.

    Key product characteristics:

    • Organotypic cell composition with primary human hepatocytes and Kupffer cells
    • Culture life time of more than 5 weeks
    • Superior liver-specific functionality as compared to 2D culture
    • Liver cytoarchitecture is maintained including full expression of bile canaliculi

    3D InSight™ Human Liver Microtissues of InSphero

    BSEP-staining of a InSphero 3D InSight™ Human Liver Microtissue showing polarized transporter protein expression and bile canaliculi.

    Morphology of human liver microtissues

    • Liver-like cytoarchitecture and stability is confirmed by H&E-staining over 28 days in culture

    3D Human Liver Microtissues culitvated over 4 weeks

    Hematoxylin and Eosin (H&E) stain of FFPE sections from 3D InSight™ Human Liver Microtissues cultivated over 4 weeks. Typical features of native human liver are apparent: cuboidal cell shape, polarization of hepatocytes and tight cellular interactions.

    • Organotypic incorporation of Kupffer cells over 28 days in culture is shown by CD68-Immunohistochemistry

    Human Liver Model with Kupffer cells remaining over 4 weeks in culture

    CD68-IHC of FFPE sections from 3D InSight™ Human Liver Microtissues cultivated over 4 weeks. Kupffer-cells are stained in deep brown. Kupffer cells remain incorporated over more than 4 weeks in culture.

    • Polarized expression of bile salt export pump (BSEP) in 3D InSight™ Human Liver Microtissues is comparable to native human liver

    Native human liver and 3D microtissue in comparison

    BSEP-Immunohistochemistry staining of FFPE sections of healthy native human liver and 3D InSight™ Human Liver Microtissues. BSEP is stained in deep brown and is indicative of a functional bile-canalicular network.

    Viability and functionality of human liver microtissues

    • Viability of 3D InSight™ Human Liver Microtissues is maintained for more than 35 days in culture as shown by ATP-content

    Viability of 3D Human Liver Microtissues more than 35 days

    Intracellular measurement of ATP-content (CellTiter-Glo®, Promega) of 3D InSight™ Human Liver Microtissues over 5 weeks in culture (n=3). The ATP-content of 3D InSight™ Human Liver Microtissues stays stable over several weeks in culture.

    • Hepatocyte specific functionality of 3D InSight™ Human Liver Microtissues for more than 35 days in culture as shown by albumin secretion

     Hepatocyte specific functionality of 3D Human Liver Models over 35 days

    Quantification of albumin released into the supernatant by 3D InSight™ Human Liver Microtissues (Albumin ELISA, Bethyl-laboratories). The secretion rate is close to native human liver (approx. 3 pg/cell*h; from Meng, 2010, Expert Opin. Drug Metab. Toxicol.).

    • Superior CYP3A4 enzyme activity of 3D InSight™ Human Liver Microtissues as compared to 2D cell culture

    CYP3A4 enzyme activity of 3D Human Liver Microtissues

    Measurement of CYP3A4-mediated midazolam hydroxylation of 3D InSight™ Human Liver Microtissues at culture day 7, 14, 21 and 28. The same hepatocyte lot was used for assessment of CYP3A4 activity in standard 2D-culture on day 2 after plating. The CYP3A4 enzyme increases over the culture period.

  • Mechanistic toxicology

    • «biased» discovery of MOA

    Long term and chronic toxicity
    Idiosyncratic toxicity
    Mitochondrial toxicity
    Oxidative stress
    Steatosis/ Phospholipidosis
    Cholestasis
    Apoptosis
    Kupffer-cell activation
    Kupffer-cell phagocytosis
    Bile-acid transporter inhibition

    • «unbiased» discovery of MOA

    Global proteomic profiling
    Transcriptomic profiling
    Histopathological evaluation

    DMPK

    CYP-induction/inhibition (DDI)
    Metabolic ID
    Low clearance compounds

    Liver disease modelling

    Diabetes
    Fibrosis
    Steatosis
    Cholestasis
    Viral infections (e.g. HCR/HBV, AAV)

  • Toxicity mechanism Cellular indicator

    Hepatotoxicity

    ATP-content
    LDH (released/intracellular)
    AST/ALT (released/intracellular)
    α-GST (released/intracellular)
    Albumin secretion
    Urea secretion
    Kupffer-cell function/toxicity IL-6 secretion
    TNF-α ELISA
    Mitochondrial impairment Mitochondrial activity
    Reactive metabolites Lower GSH content
    Metabolic competence Cytochrome activation/ induction
    Apoptosis Caspase 3/7 activation
    Steatosis, phospholipidosis Lipid accumulation
    Cholestasis Transporter proteins
    Bile-acid secretion
    Cancerogenicity DNA-synthesis
    Unbiased discovery of MOA Transcriptomics
    Proteomics
  • Catalog # Description Price in USD
    MT-02-002-01 3D InSight™ Human Liver Microtissues from primary hepatocytes (96x) 1,750
    MT-02-002-04 3D InSight™ Human Liver Microtissues from primary hepatocytes, co-culture with non-parenchymal cells (96x) 1,930
    CS-07-001-01 3D human liver microtissue maintenance medium LiMM (500 mL) 375
    Related
    Assay Kits
       
    AK-01-001-01 3D InSight™ Cytotoxicity Assay Kit 210

    For up-to-date information about our next production and shipping dates, please see our calendar here

    For volume discounts (starting at 5 plates) and shipping cost, please contact our sales staff to request a quote.

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