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ORIGINAL RESEARCH article

Front. Cell Dev. Biol.

Sec. Stem Cell Research

Volume 13 - 2025 | doi: 10.3389/fcell.2025.1528991

Aspects of zone-like identity and holotomographic tracking of human stem cell-derived liver sinusoidal endothelial cells

Provisionally accepted
Mikel Amirola MartinezMikel Amirola Martinez1Thomas CombriatThomas Combriat1,2Ingrid WilhelmsenIngrid Wilhelmsen1,3Andrea Dalmao-FernandezAndrea Dalmao-Fernandez1,4Justyna StokowiecJustyna Stokowiec1Aleksandra AizenshtadtAleksandra Aizenshtadt1,3*Stefan KraussStefan Krauss1,3
  • 1Hybrid Technology Hub, Faculty of Medicine, University of Oslo, Oslo, Norway
  • 2Njord Centre, Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Oslo, Norway
  • 3Department of Immunology and Transfusion Medicine, Oslo University Hospital, Oslo, Norway
  • 4Section for Pharmacology and Pharmaceutical Biosciences, Department of Pharmacy, Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Oslo, Norway

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Liver sinusoidal endothelial cells (LSEC) are specialized endothelial cells with unique metabolic and barrier functions adapted to the needs of the liver sinusoid. LSECs are highly sensitive to their environment, and this fragile nature causes challenges in analyzing their phenotype under in vitro conditions. In this work, we first differentiated LSEC-like cells (scLSECs) from two human pluripotent stem cell lines and characterized them by a panel of qPCR markers, immunohistochemistry, substrate oxidation for energy metabolism, scavenger function, and nitric oxide secretion. We then introduced holotomography, a technique that allows to recover quantitative and three-dimensional information about the refractive indexes of cell components, as a tool to image and track scLSEC in vitro in a minimally intrusive, label-free manner. This allowed us to describe and monitor changes in intracellular pore-like structures over time. To analyze the imaging data, we developed a machine learning-based algorithm trained to identify and then characterize these pore-like structures. Finally, we tested the possibility of inducing aspects of zone-specific LSEC phenotype and metabolism using culture-based treatments, which resulted in modest shifts in marker expression and metabolic activity. The presented strategy provides an advanced tool kit for investigating liver endothelial cells.

Keywords: LSEC, fenestration, Holotomography (HT), Pluripotent stem cells (PSC), Liver zonation

Received: 28 Nov 2024; Accepted: 03 Sep 2025.

Copyright: © 2025 Amirola Martinez, Combriat, Wilhelmsen, Dalmao-Fernandez, Stokowiec, Aizenshtadt and Krauss. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Aleksandra Aizenshtadt, Hybrid Technology Hub, Faculty of Medicine, University of Oslo, Oslo, 0372, Norway

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Supplementary Material