Comparison of iPSC-derived liver organoids and other liver cell models using proteomic analysis and CYP450 assays

Authors

• S Yang
• J Hao
• M Xia

Abstract

Hepatotoxicity induced by drugs is a primary reason for drug withdrawal from the market. A high-throughput screening method using an in vitro liver model is essential for early-stage liver toxicity assessment. Traditionally, monolayer human hepatocytes or immortalized liver cell lines (e.g., HepG2, HepaRG) have been used to evaluate liver toxicity. However, these monolayer-cultured liver cells, while useful for short-term toxicological testing, do not adequately mimic in vivo conditions and may fall short in diagnosing chronic and recurrent drug-induced liver damage. Recently, various three-dimensional (3D) liver models have been developed. This study compared iPSC-derived human liver organoids and 3D cultures of different human liver cell lines (human primary hepatocytes, HepaRG, and HepG2) with their 2D counterparts. Proteomics results indicated that iPSC-derived liver organoids matured with increased differentiation time, with their global protein expression pattern closely resembling functional liver cells at later stages. The 3D cultures of HepaRG cells exhibited higher expression of hepatic drug metabolic enzymes, such as the CYP450 enzyme family (e.g., CYP3A4), compared to their monolayer cultures, indicating enhanced physiological function. In cytochrome P450 (CYP) activity assays, human primary hepatocytes initially showed the highest CYP3A4 activity, but this activity rapidly declined over time. Conversely, iPSC-derived human liver organoids maintained sustained CYP3A4 activity and exhibited similar IC50s (0.3–0.4 μM) to HepaRG cells when treated with ketoconazole, demonstrating comparable CYP activity in vitro. Our findings suggest that iPSC-derived liver organoids represent a promising in vitro liver model for toxicity testing in drug development and the identification of potential environmental hazards.

Scientific Focus Area: Stem Cell Biology

This page was last updated on Tuesday, August 6, 2024