Unraveling MASH: Kupffer Cell-Driven Inflammation and Fibrosis within Hepatic Spheroids as a 3D Disease Model

Background: Chronic inflammation and fibrosis are central to the progression of metabolic associated steatohepatitis (MASH). Hepatic spheroids, composed of hepatocytes, stellate cells, Kupffer cells, and endothelial cells, offer a more accurate model of liver disease, capturing cell interactions and fibrosis better than traditional 2D cultures.

Materials and Methods: 3D liver spheroids were generated using HepaRG cells, stellate cells, and endothelial cells, with and without Kupffer cells, to simulate different liver conditions. Steatotic conditions were induced with palmitic and oleic acids, while MASH-like conditions were created by adding lipopolysaccharide (LPS). Cytotoxicity was measured using an LDH assay, and fibrosis and steatosis were evaluated through collagen III, fibronectin, Sirius red, and Nile red staining. Inflammatory cytokine levels (IL-6, IL-1β, TNF-α) were measured using ELISA and PCR.

Results: MASH spheroids containing Kupffer cells exhibited significantly higher levels of IL-6, IL-1β, and TNF-α, as well as increased LDH release, compared to spheroids without Kupffer cells and those in steatotic conditions. These findings suggest enhanced inflammation and cytotoxicity in MASH spheroids, highlighting the crucial role of Kupffer cells in driving inflammatory responses.

Conclusion: The 3D liver spheroid model is a valuable tool for studying MASH pathogenesis, especially for investigating liver inflammation and fibrosis. Kupffer cells are crucial for accurately modeling the disease, as they significantly amplify inflammatory and cytotoxic responses. Their inclusion is essential for understanding liver injury mechanisms, making this model effective for testing therapeutic interventions.

Publication History

Article published online:
20 January 2025