Optimized extended PCLS culture system extends tissue viability up to two weeks

Technical refinements enable prolonged tissue viability

The investigators processed 25 explanted human livers from diverse etiologies, including alcohol-associated liver disease, metabolic dysfunction-associated steatohepatitis (MASH), primary sclerosing cholangitis (PSC), hepatitis B virus (HBV)-related disease, and non-fibrotic controls.

Several experimental innovations contributed to the improved culture performance. These included compression-assisted vibratome slicing to generate more uniform tissue sections, hyperoxygenation recovery after slicing, and optimized oxygen and nutrient delivery during culture. Together, these refinements substantially improved reproducibility and extended tissue survival.

Importantly, the study also introduced a multidimensional viability assessment framework. Instead of relying solely on destructive ATP assays, the authors combined whole-plate imaging, live/dead whole-mount staining, and Seahorse metabolic analysis to dynamically monitor tissue health over time.

Dynamic remodeling reveals disease-relevant biology

A key finding of the study is that extended PCLS culture is not simply maintaining static tissue architecture. Rather, the cultured liver slices undergo active multicellular remodeling that mirrors important pathological processes observed in chronic liver disease.

Notably, cirrhotic liver-derived PCLS exhibited increasing KRT19-positive ductular cells during culture, consistent with ductular reaction and epithelial plasticity. In contrast, healthy liver slices showed evidence of hepatocyte regenerative clusters. These findings suggest that the model can simultaneously capture regeneration, fibrosis, and biliary remodeling within intact human liver tissue.

Translational implications for liver disease research

The study highlights the growing importance of human-derived New Approach Methodologies (NAMs) in biomedical research. Because PCLS retains native immune cells, stromal components, and spatial tissue organization, it may provide advantages over conventional cell culture systems and some animal models.

The platform could support multiple translational applications, including anti-fibrotic drug testing, investigation of hepatocyte plasticity, precision medicine approaches, and validation of therapeutic targets in human liver tissue. As spatial omics and single-cell technologies continue to evolve, extended PCLS culture may become an increasingly valuable bridge between mechanistic discovery and clinical translation.

Source:

First Hospital of Jilin University

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