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

STAR Protoc. 2024 Sep 20;5(3):103234. doi: 10.1016/j.xpro.2024.103234. Epub 2024 Aug 10.

Abstract

To improve human hepatotoxicity prediction, in vitro liver cell models replicating hepatocyte function, drug metabolism, and toxicity are required. Here, we present a protocol for creating 3D primary human hepatocyte (PHH) cell models using the RASTRUM Platform. We describe the process for PHH model generation; procedures for characterizing the PHH model, including viability, albumin production, and CYP450 inducibility; and drug treatment using acetaminophen and troglitazone. This protocol has applications in upscaling phenotypic hepatotoxicity applications.

Keywords: Biotechnology and bioengineering; Cell Biology; Cell culture; Molecular Biology.

MeSH terms

  • Acetaminophen / pharmacology
  • Cell Culture Techniques / methods
  • Cell Culture Techniques, Three Dimensional / methods
  • Cell Survival / drug effects
  • Cells, Cultured
  • Hepatocytes* / cytology
  • Hepatocytes* / drug effects
  • Hepatocytes* / metabolism
  • Humans
  • Phenotype
  • Primary Cell Culture / methods
  • Troglitazone / pharmacology

Substances

  • Troglitazone
  • Acetaminophen