# From Patient Liver Tissue to Organoids: Establishment of a Translational Platform Using Healthy, Steatotic, and Cirrhotic Tissue Sources

## TL;DR

Researchers developed liver organoids from patient tissue samples, including healthy, fatty, and cirrhotic livers, to better study liver diseases and test treatments.

## Contribution

A high-success-rate method for generating liver organoids from diverse patient tissues, enabling disease modeling and drug testing.

## Key findings

- 45 patient-derived liver organoids were successfully established with an 82% initiation success rate.
- Organoids showed high proliferation and co-expression of hepatocyte and cholangiocyte markers.
- Mathematical modeling revealed a biphasic growth pattern in the organoids.

## Abstract

What are the main findings?
45 patient-derived liver organoids from healthy, steatotic, and cirrhotic tissue were successfully established with an 82% initiation success rate after optimizing the Broutier protocol.The organoids exhibited high proliferation, co-expression of hepatocyte markers (Albumin and HNF4α) and cholangiocyte markers (CK19), sporadic LGR5 positivity, and a biphasic growth pattern, as revealed by mathematical modeling.

45 patient-derived liver organoids from healthy, steatotic, and cirrhotic tissue were successfully established with an 82% initiation success rate after optimizing the Broutier protocol.

The organoids exhibited high proliferation, co-expression of hepatocyte markers (Albumin and HNF4α) and cholangiocyte markers (CK19), sporadic LGR5 positivity, and a biphasic growth pattern, as revealed by mathematical modeling.

What are the implications of the main findings?
These organoid cultures provide a physiologically relevant 3D platform for dissecting the mechanisms of MASLD progression and comparing disease phenotypes across patient cohorts, overcoming the limitations of traditional 2D cultures.The combination of molecular profiling and quantitative growth modeling paves the way for precision drug testing, integration of systems biology, and personalized therapeutic strategies in liver disease research.

These organoid cultures provide a physiologically relevant 3D platform for dissecting the mechanisms of MASLD progression and comparing disease phenotypes across patient cohorts, overcoming the limitations of traditional 2D cultures.

The combination of molecular profiling and quantitative growth modeling paves the way for precision drug testing, integration of systems biology, and personalized therapeutic strategies in liver disease research.

Metabolic dysfunction-associated steatotic liver disease (MASLD) and its consequences represent a growing global health burden that urgently requires physiologically relevant in vitro models beyond conventional 2D culture systems. In this study, we report the successful establishment of 45 patient-derived liver organoid lines. These organoids were generated from healthy, steatotic and cirrhotic tissues collected from 207 liver surgeries at RWTH University Hospital Aachen, with an initiation success rate of 82%. The organoids were propagated for at least six passages using an optimized protocol. Multiplex immunofluorescence analysis revealed highly proliferative structures with approximately 40% Ki-67-positive cells expressing hepatocyte (Albumin and HNF4α) and cholangiocyte (CK19) markers. Intermittent LGR5 staining suggested the presence of liver progenitor cell features. Quantitative PCR results confirmed variable HNF4α expression, indicating inter-patient heterogeneity in differentiation status. Time-lapse imaging combined with mathematical modeling uncovered a biphasic growth dynamic with an initial linear expansion in the first 15 h, followed by exponential growth (doubling time ≈ 20.6 h) between 30 and 72 h. Overall, our workflow produced genetically and phenotypically stable liver organoids that recapitulate essential features of various hepatic conditions. This provides a solid foundation for disease modeling, potential drug testing, and quantitative systems biology.