Biology subjects

Koning, J. J.

Publications and source records attributed to Koning, J. J..

4 recordsLinked to original sources

bioRxiv2025

Integration of lymphatic vasculature to a human lymph node-on-chip enhances physiological immune properties

To study systemic human innate and adaptive immune responses in detail, competent in vitro lymph node (LN) models with LN stromal cells (LNSCs) are required to recapitulate the physiological microenvironment. The multicellular organisation of LNs possesses a challenge for designing such microphysiological systems (MPS), particularly with the structural complexity of LNs and the lymphatic vasculature. Here, we established an organotypic LN model with integrated lymphatics in an organ-on-chip (OoC) platform containing a printed sacrificial structure, and studied the influence of a perfused lymphatic endothelial cell (LEC)-lined channel on the LN-on-chip microenvironment. Upon one-week of culture under lymphatic flow, LECs lined the tubular structure forming a lymphatic vessel through the LN model, and stable metabolic conditions within the LN-on-chip were confirmed. Interestingly, LECs in the LN-on-chip displayed the phenotype found in human LNs with upregulation of LEC-specific LN markers, such as atypical chemokine receptor 4 (ACKR4). The presence of the LEC-lined perfused vessel in the LN-on-chip resulted in the increase of native immune cells, most notably B cells, and the secretion of survival and migratory signals, namely interleukin-7 (IL-7) and CC motif chemokine ligand 21 (CCL21). Likewise, LECs promoted the abundance of immune cell clusters closer to the vessel. As such, these features represent an enhanced physiological microenvironment to allow for immune cell migration and interactions for efficient LN functioning. This approach paves the way for LN integration into multi-OoC (MOC) platforms to investigate immunological crosstalk between tissue-derived factors, immune cell trafficking and organ-specific adaptive immune responses.

bioengineering↗

bioRxiv2025

A Sacrificial 3D Printed Vessel-on-Chip Demonstrates a Versatile Approach to Model Connective Tissue Pathology

For in vitro organ models, perfused vasculature is crucial to overcome nutrient diffusion limits and to generate immunocompetent models by allowing trans-endothelial migration of immune cells in and out of the tissue. However, vasculature is often disregarded due to its complexity to generate and the necessity to integrate flow. The aim here was to overcome these limitations by combining 3D printing and multi-organ-chip technology to generate a vascularized, fibroblast-populated connective tissue matrix on-chip. A 3D printed, sacrificial, water-dissolvable structure was incorporated into a multi-organ-chip to generate hollow channels within a collagen/fibrin hydrogel. Subsequently, the channels were populated with endothelial cells. Different hydrogel concentrations of fibrin were used to mimic healthy and early granulation tissue. The vessels were perfused, and stable metabolic/viability conditions (lactate dehydrogenase, glucose, lactate) acquired after 3 days for 7 days total. In high fibrin gels, angiogenic sprouting and increased secretion of angiogenic cytokines was observed. Perfusion with monocytes revealed differentiation into macrophages and migration across the endothelium into the tissue. In conclusion, the versatile, easy method to pattern hydrogels in multi-organ-chips can serve as the basis to build the next generation of vascularized, immunocompetent human organ models, and opens new possibilities to study health and disease.

bioengineering↗

bioRxiv2020

Development of follicular dendritic cells in lymph nodes depends on retinoic acid mediated signaling

Specialized stromal cells occupy and help define B- and T cell domains, which is crucial for proper functioning of our immune system. Signaling through lymphotoxin and TNF-receptors is crucial for development of different stromal subsets which are thought to arise from a common precursor. However, mechanisms that control the selective generation of the different stromal phenotypes are not known. Here we show that in mice, retinoic acid mediated signaling is important for the differentiation of precursors towards the Cxcl13pos follicular dendritic cell (FDC) lineage, while blocking lymphotoxin mediated Ccl19pos fibroblastic reticular cell (FRC) lineage differentiation. Consequently, we see at day of birth Cxcl13posCcl19neg/low and Cxcl13neg/lowCcl19pos cells within neonatal lymph nodes. Furthermore, ablation of retinoic acid receptor signaling in stromal precursors early after birth reduces Cxcl13 expression, while in addition, complete blockade of retinoic acid signaling prevents formation of FDC networks in lymph nodes.

immunology↗

bioRxiv2020

A molecular map of lymph node blood vascular endothelium at single cell resolution

Blood vascular endothelial cells (BECs) control the immune response by regulating immune cell recruitment, metabolite exchange and blood flow in lymphoid tissues. However, the diversity of BEC and their origins during immune angiogenesis remain poorly understood. Here we profile transcriptomes of BEC from mouse peripheral lymph nodes and map key phenotypes to the vasculature. Our analysis identifies multiple novel subsets including a venous population whose gene signature predicts an unexpectedly selective role in myeloid cell (vs lymphocyte) recruitment to the medulla, confirmed by 2 photon videomicroscopy. We define five phenotypes of capillary lining BEC including a capillary resident regenerative population (CRP) that displays stem cell and migratory gene signatures and contributes to homeostatic BEC turnover and to vascular neogenesis after immunization. Trajectory analyses reveal retention of developmental programs along a progression of cellular phenotypes from CRP to mature venous and arterial BEC subsets. Overall, our single cell atlas provides a molecular blueprint of the lymph node blood vasculature and defines subset specialization for immune cell recruitment and vascular homeostasis.

immunology↗