The fibrotic response of primary liver spheroids recapitulates in vivo hepatic stellate cell activation
| Autor: | Sofia B. Leite, Ayla Smout, Inge Mannaerts, Olivier Frey, Stefaan Verhulst, Elise Anne van Os, Nathalie Eysackers, Leo A. van Grunsven, Andreas Hierlemann, T. Roosens |
|---|---|
| Přispěvatelé: | Basic (bio-) Medical Sciences, Liver Cell Biology, Faculty of Medicine and Pharmacy, Translational Liver Cell Biology |
| Rok vydání: | 2020 |
| Předmět: |
Liver Cirrhosis
stellate cells Biophysics Bioengineering 02 engineering and technology Toxicology Biomaterials bioreactor 03 medical and health sciences Mice Fibrosis In vivo medicine Hepatic Stellate Cells Animals acetaminophen liver fibrosis 030304 developmental biology Medicine(all) 0303 health sciences Chemistry Spheroid Cell Biology 021001 nanoscience & nanotechnology medicine.disease Hepatic stellate cell activation In vitro Cell biology medicine.anatomical_structure Liver Mechanics of Materials Hepatocyte Liver spheroids Ceramics and Composites Hepatic stellate cell Hepatocytes Hepatocyte dedifferentiation 0210 nano-technology |
| Zdroj: | Biomaterials. 261 |
| ISSN: | 1878-5905 |
| Popis: | A major obstacle in the development of efficient therapies for progressive liver fibrosis is the lack of representative in vitro models of liver fibrosis to aid in understanding the mechanisms of the disease and to promote the development of pharmaceuticals. Our aim was to develop a relevant in vitro mouse liver fibrosis model, based on the central hypothesis that liver fibrosis in vitro cannot be studied using only hepatic stellate cells (HSCs)-the main producer of scar tissue during fibrosis-, but requires cultures in which at least hepatocytes are integrated. We established robust methods to generate co-culture spheroids from freshly isolated mouse hepatocytes and HSCs. Characteristics and functionality of these spheroids were analyzed by qPCR of cell-type specific markers, CYP induction and immunohistochemistry. Compound toxicity was determined by ATP-assays. Hepatocytes and HSCs maintained their cell-type specific marker expression over a 15-day culture period without major hepatocyte dedifferentiation or HSC activation. Exposure of spheroids to TGFβ can directly activate HSCs, while acetaminophen exposure mounts a hepatocyte damage dependent activation of HSCs. Pharmaceuticals with known anti-fibrotic properties, such as Valproic acid and Verteporfin, reduce HSC activation in response to hepatocyte damage in these cultures. A comparison between the fibrotic response of the spheroid co-cultures and in vivo activated HSCs showed that these 3D co-cultures are more representative than the commonly used 2D HSC monocultures. Finally, we showed that the 3D cultures can be integrated in microfluidic chips. We conclude that our hepatocyte-stellate cell-spheroid cultures are a robust in vitro model of liver fibrosis. This model could be used to further unravel the mechanism of HSC activation and facilitate the discovery of, or testing for novel anti-fibrotic compounds, as these spheroids better reproduce HSC in vivo activation compared to the more traditional 2D mono-culture models. |
| Databáze: | OpenAIRE |
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