Primary Hepatic 3D culture systems

In the past decade 3D liver cultures as developed in leaps and bounds for study of liver functions, and disease progression. Most of these culture systems are also adopted commercially, for testing drug toxicity, drug interactions and pharmacokinetics. Such advances stems from the inability of traditional monolayer cultures at proving an accurate representation of liver cells and subsequent function, thus yielding results with poor predictability (1,2).

3D culture systems for primary human hepatocyte cultures (PHH)

PHH Spheroids are 3D aggregates of approximately 200-300μm. Spheroid cultures are particularly effective at producing mature hepatocytes, resulting in long-term viability and stabile liver functions. Several methods have been developed thus far to produce liver spheroids namely, Stirring bioreactors, Hanging drop aggregates, or culturing on low attachment cell repellant plates (3). The latter method is currently the go-to choice among the spheroid formation platforms, due to its reliability and reproducibility (4). Each system comprises of different techniques in spheroid formation, and also differs in its applications.

Applications of 3D PHH cultures

The rapid development of 3D culture systems for liver cells has had an equal effect on the production of translatable models for mimicking drug response, and toxicity along with physiologically relevant liver models for studying pathophysiology of liver diseases. This correlative effect has in turn led to increased predictability in primary human cultures for analyzing chronic effects of drug toxicity, and metabolism of low clearance drugs. Furthermore, these innovative culture platforms also enable drug screens for treatment of liver disease such as non-alcoholic fatty liver disease (NAFLD), hepatitis and liver cancer. As an added benefit, majority of these cultures can be adapted to high throughout or high content research systems. However standardization of these systems to ensure representative hepatic phenotype as well, in-vivo like functional properties still needs careful optimization. With ongoing research into novel 3D culture platforms, hepatic 3D systems will continue to grow in its value in future drug development, as well as understanding the mechanisms of liver disease and potential treatments (3,4).


1. V. M. Lauschke, S. U. Vorrink, S. M. L. Moro, F. Rezayee, Å. Nordling, D. F. G. Hendriks, C. C. Bell, R. Sison‐Young, B. K. Park, C. E. Goldring, E. Ellis, I. Johansson, S. Mkrtchian, T. B. Andersson, M. Ingelman‐Sundberg, Hepatology 2016, 64, 1743.

2. C. C. Bell, V. M. Lauschke, S. U. Vorrink, H. Palmgren, R. Duffin, T. B. Andersson, M. Ingelman‐Sundberg, Drug Metab. Dispos. 2017, 45, 419.

3. Lauschke VM, Shafagh RZ, Hendriks DFG, Ingelman-Sundberg M. 3D Primary Hepatocyte Culture Systems for Analyses of Liver Diseases, Drug Metabolism, and Toxicity: Emerging Culture Paradigms and Applications. Biotechnol J. 2019 Jul;14(7):e1800347.

4. Raic, A., Oberfrank, C., Birk, B. et al. 3D-Hunde-Lebersphäroide für die präklinische in vitro-Testung. Biospektrum 27, 736–738 (2021).