{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:17:51Z","timestamp":1790792271178,"version":"4.1.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T00:00:00Z","timestamp":1690243200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005416","name":"Norges Forskningsr\u00e5d","doi-asserted-by":"publisher","award":["262613"],"award-info":[{"award-number":["262613"]}],"id":[{"id":"10.13039\/501100005416","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006095","name":"Helse S\u00f8r-\u00d8st RHF","doi-asserted-by":"publisher","award":["2021068"],"award-info":[{"award-number":["2021068"]}],"id":[{"id":"10.13039\/501100006095","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Bioeng. Biotechnol."],"abstract":"<jats:p><jats:bold>Background:<\/jats:bold> There is a significant need for predictive and stable <jats:italic>in vitro<\/jats:italic> human liver representations for disease modeling and drug testing. Hepatic stellate cells (HSCs) and liver sinusoidal endothelial cells (LSECs) are important non-parenchymal cell components of the liver and are hence of relevance in a variety of disease models, including hepatic fibrosis. Pluripotent stem cell- (PSC-) derived HSCs (scHSCs) and LSECs (scLSECs) offer an attractive alternative to primary human material; yet, the suitability of scHSCs and scLSECs for extended <jats:italic>in vitro<\/jats:italic> modeling has not been characterized.<\/jats:p><jats:p><jats:bold>Methods:<\/jats:bold> In this study, we describe the phenotypic and functional development of scHSCs and scLSECs during 14 days of 2D <jats:italic>in vitro<\/jats:italic> culture. Cell-specific phenotypes were evaluated by cell morphology, immunofluorescence, and gene- and protein expression. Functionality was assessed in scHSCs by their capacity for intracellular storage of vitamin A and response to pro-fibrotic stimuli induced by TGF-\u03b2. scLSECs were evaluated by nitric oxide- and factor VIII secretion as well as endocytic uptake of bioparticles and acetylated low-density lipoprotein. Notch pathway inhibition and co-culturing scHSCs and scLSECs were separately tested as options for enhancing long-term stability and maturation of the cells.<\/jats:p><jats:p><jats:bold>Results and Conclusion:<\/jats:bold> Both scHSCs and scLSECs exhibited a post-differentiation cell type-specific phenotype and functionality but deteriorated during extended culture with PSC line-dependent variability. Therefore, the choice of PSC line and experimental timeframe is crucial when designing <jats:italic>in vitro<\/jats:italic> platforms involving scHSCs and scLSECs. Notch inhibition modestly improved long-term monoculture in a cell line-dependent manner, while co-culturing scHSCs and scLSECs provides a strategy to enhance phenotypic and functional stability.<\/jats:p>","DOI":"10.3389\/fbioe.2023.1223737","type":"journal-article","created":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T18:12:55Z","timestamp":1690308775000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Characterization of human stem cell-derived hepatic stellate cells and liver sinusoidal endothelial cells during extended in vitro culture"],"prefix":"10.3389","volume":"11","author":[{"given":"Ingrid","family":"Wilhelmsen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikel","family":"Amirola Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justyna","family":"Stokowiec","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chencheng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandra","family":"Aizenshtadt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Krauss","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,7,25]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1016\/j.lfs.2017.11.018","article-title":"TGF-\u03b21 signaling regulates mouse hepatic stellate cell differentiation via the Jagged1\/Notch pathway","volume":"192","author":"Aimaiti","year":"2018","journal-title":"Life Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"881","DOI":"10.1007\/s10616-019-00329-y","article-title":"TGF-\u03b21 signaling activates hepatic stellate cells through Notch pathway","volume":"71","author":"Aimaiti","year":"2019","journal-title":"Cytotechnology"},{"key":"B3","doi-asserted-by":"publisher","first-page":"998","DOI":"10.1002\/hep.22721","article-title":"Mesenchymal origin of hepatic stellate cells, submesothelial cells, and perivascular mesenchymal cells during mouse liver development","volume":"49","author":"Asahina","year":"2009","journal-title":"Hepatology"},{"key":"B4","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.jhep.2018.09.014","article-title":"Burden of liver diseases in the world","volume":"70","author":"Asrani","year":"2019","journal-title":"J. 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