Biology subjects

van Grinsven, E. J.

Publications and source records attributed to van Grinsven, E. J..

3 recordsLinked to original sources

bioRxiv2025

Immortalization of human nasal and bronchial airway epithelial cells for genome editing applications

In vitro air-liquid interface culture of airway epithelial cells is used as a model system to study respiratory diseases. This culture system not only overcomes the need for animal models or continuous biopsies from individuals but also enables studies of pathophysiology associated with the disease in a patient background. Human airway basal cells serve as progenitor cells for a functional pseudostratified airway epithelium composed mainly of multiciliated and secretory cells. However, due to the limited ability of basal cells to proliferate and differentiate, the long-term use of primary material in culture is restricted. This challenges research that requires genome editing. Here, we describe airway stem cells from nasal and bronchial origin immortalized by hTERT overexpression followed by polyclonal expansion. We demonstrate that this diverse panel of cell lines shows differentiation patterns similar to primary stem cells and can be used for lentiviral and CRISPR/Cas9 genome editing. These cell lines and optimized protocols facilitate airway biology research and disease phenotyping.

cell biology↗

bioRxiv2025

Microtubule organization and molecular architecture of ciliary basal bodies in multiciliated airway cells

Microtubule organization depends on cell type and function. Microtubule networks of many differentiated cell types, such as epithelial cells, are poorly understood due to their high density. Here, we used expansion microscopy to quantitatively describe the three-dimensional organization of the microtubule network in human airway multiciliated cells. In these cells, most apical and apicobasal microtubules nucleate and anchor at the basal foot, a part of the ciliary basal body. Using a newly developed volumetric averaging tool, we generated a high-resolution 3D map of the basal body, delineating the position of structural components and proteins involved in microtubule nucleation and anchoring. {gamma}-TuRC, its binding partners NEDD1 and Augmin/HAUS, and centriolar appendage proteins ninein and AKNA localize to the basal foot. Functional analyses demonstrated that NEDD1, but not ninein or HAUS are essential for basal foot-dependent microtubule organization. Our data reveal the distinct architecture of microtubule-organizing centers responsible for the formation of dense microtubule arrays in multiciliated cells.

cell biology↗

bioRxiv2021

Optical nanoscopy reveals SARS-CoV-2-induced remodeling of human airway cells

A better understanding of host cell remodeling by the coronavirus SARS-CoV-2 is urgently needed to understand viral pathogenesis and guide drug development. Expression profiling and electron microscopy have frequently been used to study virus-host interactions, but these techniques do not readily enable spatial, sub-cellular and molecular analysis of specific cellular compartments. Here, we use diffraction-unlimited fluorescence microscopy to analyze how SARS-CoV-2 infection exploits and repurposes the subcellular architecture of primary human airway cells. Using STED nanoscopy, we detect viral entry factors along the motile cilia of ciliated cells and visualize key aspects of the viral life cycle. Using Tenfold Robust Expansion (TREx) microscopy, we analyze the extensively remodeled three-dimensional ultrastructure of SARS-CoV-2-infected ciliated cells and uncover Golgi fragmentation, emergence of large and atypical multivesicular bodies enclosing viral proteins, ciliary clustering, and remodeling of the apical surface. These results demonstrate a broadly applicable strategy to study how viruses reorganize host cells with spatial and molecular specificity and provide new insights into SARS-CoV-2 infection in primary human cell models.

cell biology↗