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Goulart E , de Caires-Junior LC , Telles-Silva KA , Araujo BHS , Kobayashi GS , Musso CM , Assoni AF , Oliveira D , Caldini E , Gerstenhaber JA , Raia S , Lelkes PI , Zatz M
Adult and iPS-derived non-parenchymal cells regulate liver organoid development through differential modulation of Wnt and TGF-beta
Stem Cell Res Ther. 2019 Aug 15;10(1) :258
PMID: 31416480    PMCID: PMC6694663   
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Abstract
BACKGROUND: Liver organoid technology holds great promises to be used in large-scale population-based drug screening and in future regenerative medicine strategies. Recently, some studies reported robust protocols for generating isogenic liver organoids using liver parenchymal and non-parenchymal cells derived from induced pluripotent stem cells (iPS) or using isogenic adult primary non-parenchymal cells. However, the use of whole iPS-derived cells could represent great challenges for a translational perspective. METHODS: Here, we evaluated the influence of isogenic versus heterogenic non-parenchymal cells, using iPS-derived or adult primary cell lines, in the liver organoid development. We tested four groups comprised of all different combinations of non-parenchymal cells for the liver functionality in vitro. Gene expression and protein secretion of important hepatic function markers were evaluated. Additionally, liver development-associated signaling pathways were tested. Finally, organoid label-free proteomic analysis and non-parenchymal cell secretome were performed in all groups at day 12. RESULTS: We show that liver organoids generated using primary mesenchymal stromal cells and iPS-derived endothelial cells expressed and produced significantly more albumin and showed increased expression of CYP1A1, CYP1A2, and TDO2 while presented reduced TGF-beta and Wnt signaling activity. Proteomics analysis revealed that major shifts in protein expression induced by this specific combination of non-parenchymal cells are related to integrin profile and TGF-beta/Wnt signaling activity. CONCLUSION: Aiming the translation of this technology bench-to-bedside, this work highlights the role of important developmental pathways that are modulated by non-parenchymal cells enhancing the liver organoid maturation.
Notes
1757-6512 Goulart, Ernesto de Caires-Junior, Luiz Carlos Telles-Silva, Kayque Alves Araujo, Bruno Henrique Silva Kobayashi, Gerson S Musso, Camila Manso Assoni, Amanda Faria Oliveira, Danyllo Caldini, Elia Gerstenhaber, Jonathan A Raia, Silvano Lelkes, Peter I Zatz, Mayana ORCID: http://orcid.org/0000-0003-3970-8025 2015/14821-1/fapesp 2013/08028-1/fapesp 2017/16283-2/fapesp Journal Article England Stem Cell Res Ther. 2019 Aug 15;10(1):258. doi: 10.1186/s13287-019-1367-x.