Scalable 2D and 3D human hepatocyte platforms for target validation and compound screening in early drug discovery pipelines of liver diseases

Physiologically relevant in-vitro liver systems are critical for translational research in metabolic diseases, including Metabolic Dysfunction-Associated Steatotic Liver (MASL) and Metabolic Dysfunction-Associated Steatohepatitis (MASH).  Evotec has established advanced primary human hepatocyte platforms encompassing 2D sandwich cultures and long-term 3D spheroid models capable of sustaining hepatic phenotype and metabolic competence. The 3D spheroid model also incorporates multi-cellular complexity and enables the modeling of key disease phenotypes such as steatosis, fibrotic signaling, extracellular matrix remodeling, and metabolically driven responses under defined exposure conditions. 

In this session, Nicole Otto and Dr. Ana Martinez Hernandez will present their evaluation of gene expression data generated using Evotec’s established 2D cultures and 3D hepatocyte platforms. Leveraging these mature invitro liver models, they will highlight gene expression profiling approaches to characterize hepatocyte phenotype and metabolic competence. The session will also feature insights from fibrosis induction paradigms and fatty acid–treated hepatocyte systems, illustrating how molecular readouts capture relevant responses across the platform.  

The speakers will present performance data from high throughput gene expression workflows, including direct lysis from 96-well 2D and 3D spheroid formats. Comparative results across culture formats and expression systems will identify approaches that maintain biological resolution, speed up studies, and eliminate the need for sample pooling. In addition, this work will also show Evotec’s Expression Profile Platform supports customizable gene panels (2–25+ targets) and processes more than 26,000 samples annually, enabling data-driven target identification and helping develop validation workflows for future partner programs. 

Together, these capabilities show how Evotec combines physiologically relevant liver models with scalable molecular profiling to enable mechanistic, drug metabolism, and early toxicology studies within discovery collaborations. 

Learning Objectives:

• Assess the scientific and translational advantages of 2D sandwich and long-term 3D human hepatocyte spheroid models for mechanistic modeling of metabolic liver disease. 
• Interpret gene expression–based readouts used to characterize hepatocytic differentiation, metabolic competence, and pro-fibrotic pathway activation in multi-cellular liver systems. 
• Evaluate direct lysis–based gene expression approaches, including Invitrogen™ Cells-to-Ct workflows, and their role in enabling scalable, high-throughput molecular profiling within automated discovery environments.
   

For Research Use Only. Not for use in diagnostic procedures.