AUTHOR=Amirola-Martinez Mikel , Combriat Thomas , Ferencevic Katharina , Wilhelmsen Ingrid , Dalmao-Fernandez Andrea , Olsen Petter Angell , Stokowiec Justyna , Aizenshtadt Aleksandra , Krauss Stefan 

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

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=Volume 13 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2025.1528991

DOI=10.3389/fcell.2025.1528991

ISSN=2296-634X

ABSTRACT=IntroductionLiver 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.MethodsIn 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.Results and discussionHolotomography and developed machine learning-based algorithm for image processing allowed us to describe and monitor changes in intracellular pore-like structures over time. 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.