Supplementary dataset for "Characterization of Human Stem Cell-Derived Hepatic Stellate Cells and Liver Sinusoidal Endothelial Cells During Extended in vitro Culture"https://doi.org/10.18710/4MJLC3Wilhelmsen, IngridDataverseNO2023-09-262023-09-28T21:06:00Z<p>The dataset contains confocal immunofluorescence images of human pluripotent stem cell-derived hepatic stellate cells (scHSCs) and human primary HSCs (pHSCs). </p> <p>In the homeostatic liver, HSCs exist in a quiescent state. Upon liver injury, HSCs differentiate into an activated, myofibroblast-like state. The activated HSCs are phenotypically characterized by the intracellular expression of alpha-smooth muscle actin (a-SMA). Transforming growth factor beta (TGF-b) is a potent HSC activator and is commonly used to assess the activation capacity of cultured HSCs.</p> <p>In this dataset, scHSCs and pHSCs were grown in vitro and stained with immunofluorescent labels for the HSC marker PDGFR-b as well as the activation-related proteins a-SMA and COL1a1. Image analysis of the dataset was performed to quantify a-SMA fluorescence intensity to assess the activation status of the HSCs in the presence and absence of TGF-b. </p> <p>An example script to analyze the data is provided on GitHub: https://github.com/ingridwilhelmsen/a-SMA_analysis/blob/main/a-SMA_analysis-Activation.ipynb</p>Medicine, Health and Life SciencesHepatic stellate cell (HSC)a-smooth muscle actin (a-sMA)Induced pluripotent stem cellsLiver sinusoidal endothelial cell (LSEC)Induced pluripotent stem cellConfocal immunofluorescence microscopyEnglish<p> AUTHOR=Wilhelmsen Ingrid, Amirola Martinez Mikel, Stokowiec Justyna, Wang Chencheng, Aizenshtadt Aleksandra, Krauss Stefan</p> <p> TITLE=Characterization of human stem cell-derived hepatic stellate cells and liver sinusoidal endothelial cells during extended in vitro culture </p> <p> JOURNAL=Frontiers in Bioengineering and Biotechnology </p> <p> VOLUME=11</p> <p> YEAR=2023</p> <p> URL=https://www.frontiersin.org/articles/10.3389/fbioe.2023.1223737 </p> <p> DOI=10.3389/fbioe.2023.1223737 </p> <p> ISSN=2296-4185 </p> <p> ABSTRACT=Background: There is a significant need for predictive and stable in vitro 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 in vitro modeling has not been characterized.Methods: In this study, we describe the phenotypic and functional development of scHSCs and scLSECs during 14 days of 2D in vitro 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-β. 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.Results and Conclusion: 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 in vitro 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.</p>, doi, 10.3389/fbioe.2023.1223737, https://doi.org/10.3389/fbioe.2023.12237372023-09-26Wilhelmsen, IngridKrauss, StefanAizenshtadt, AleksandraCombriat, ThomasAmirola Martinez, MikelStokowiec, JustynaWang, ChenchengWilhelmsen, Ingrid2023-07-062022-10-012022-12-15Image dataCC0 1.0