Protocol to create phenotypic primary human hepatocyte cultures using the RASTRUM 3D cell model platform.

Area of Science:

• Hepatology
• Drug Metabolism
• Toxicology

Background:

• Accurate prediction of drug-induced liver injury (DILI) is crucial for patient safety.
• Existing in vitro models often fail to fully replicate human liver function, limiting their predictive power.
• There is a need for advanced in vitro models that recapitulate key hepatocyte functions for robust toxicity testing.

Purpose of the Study:

• To present a protocol for generating 3D primary human hepatocyte (PHH) cell models.
• To establish a reliable method for characterizing PHH model functionality, including viability, albumin production, and CYP450 inducibility.
• To demonstrate the utility of these PHH models in drug-induced hepatotoxicity studies.

Main Methods:

• Utilized the RASTRUM Platform for the 3D culture of primary human hepatocytes (PHHs).
• Developed procedures for PHH model generation and subsequent characterization assays.
• Performed drug treatment experiments using known hepatotoxicants like acetaminophen and troglitazone.

Main Results:

• Successfully generated functional 3D PHH models exhibiting sustained viability.
• Demonstrated characteristic hepatocyte functions, including albumin production and inducible CYP450 enzyme activity.
• Observed distinct responses to acetaminophen and troglitazone, validating the model's capacity for toxicity assessment.

Conclusions:

• The presented protocol enables the creation of advanced 3D PHH models for in vitro hepatotoxicity prediction.
• These models offer a robust platform for evaluating drug metabolism and toxicity, improving upon traditional methods.
• This approach facilitates the upscaling of phenotypic hepatotoxicity testing, aiding in safer drug development.