On-chip human lymph node stromal network for evaluating dendritic cell and T-cell trafficking

Kwee, Brian J.; Akue, Adovi; Sung, Kyung Eun

doi:10.1101/2023.03.21.533042

Cited by 4 publications

(3 citation statements)

References 40 publications

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Section: Resultsmentioning

confidence: 99%

“…In Col/Fib gel, some of the T cells migrated into the center lane, and a small number were observed to co-localize with HLFs there when imaged after 2 days, similar to prior reports. 46,62 Interestingly, co-cultures in the GelSH + 60% collagen gels yielded HLF spreading and redistribution of the T cells in some regions but not others. Stromal cell morphology in the GelSH + 60% collagen gel was distinct and more variable than the other groups.…”

Section: Porous Photo-crosslinkable Gels Enabled Cd4+ T Cell Migratio...mentioning

confidence: 99%

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Mitigating reactive oxygen species production and increasing gel porosity improves lymphocyte motility and fibroblast spreading in photocrosslinked gelatin-thiol hydrogels

Ozulumba,

Zatorski,

Arneja

et al. 2024

Preprint

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On-chip 3D culture systems that incorporate immune cells such as lymphocytes and stromal cells are needed to model immune organs in engineered systems such as organs-on-chip. Photocrosslinking is a useful tool for creating such immune-competent hydrogel cultures with spatial cell organization. However, loss of viability and motility in photocrosslinked gels can limit its utility, especially when working with fragile primary cells. We hypothesized that optimizing photoexposure-induced ROS production, hydrogel porosity or a combination of both factors was necessary to sustain cell viability and motility during culture in photocrosslinked gelatin-thiol (GelSH) hydrogels. Jurkat T cells, primary human CD4+ T cells and human lymphatic fibroblasts were selected as representative lymphoid immune cells to test this hypothesis. Direct exposure of these cells to 385 nm light and LAP photoinitiator dramatically increased ROS levels. Pretreatment with an antioxidant, ascorbic acid (AA), protected the cells from light + LAP-induced ROS and was non-toxic at optimized doses. Furthermore, scanning electron microscopy showed that native GelSH hydrogels had limited porosity, and that adding collagen to GelSH precursor before crosslinking markedly increased gel porosity. Next, we tested the impact of AA pretreatment and increasing gel porosity, alone or in combination, on cell viability and function in 3D GelSH hydrogel cultures. Increasing gel porosity, rather than AA pretreatment, was more critical for rescuing viability of Jurkat T cells and spreading of human lymphatic fibroblasts in GelSH-based gels, but both factors improved the motility of primary human CD4+ T cells. Increased porosity enabled formation of spatially organized co-cultures of primary human CD4+ T cells and human lymphatic fibroblasts in photo-crosslinked gels in a multi-lane microfluidic chip, towards modeling the lymphoid organ microenvironment. Some optimization is still needed to improve homogeneity between regions on the chip. These findings will enable researchers utilizing photocrosslinking methods to develop immunocompetent 3D culture models that support viability and function of sensitive lymphoid cells.

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Section: Resultsmentioning

confidence: 99%

Section: Porous Photo-crosslinkable Gels Enabled Cd4+ T Cell Migratio...mentioning

confidence: 99%

Mitigating reactive oxygen species production and increasing gel porosity improves lymphocyte motility and fibroblast spreading in photocrosslinked gelatin-thiol hydrogels

Ozulumba,

Zatorski,

Arneja

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…A vital feature across all lymph node-on-chip models to date is their lack of stromal cells, in contrast to other tissue/organ models-on-chip, which all contained a stromal component, as detailed above. Currently, there is one chip model has included fibroblast reticular cells (FRCs) from a cell line source ( 75 ), which showcased DC and T cell migration towards this FRC compartment. There is clearly an unmet need for incorporation of these FRCs, given their role in not only shaping immune responses within the lymph node ( 120 ), but also their importance for immune cell survival and functioning in a 3D environment ( 121 ).…”

Section: Lymphoid Vasculature and Organsmentioning

confidence: 99%

In vitro immunity: an overview of immunocompetent organ-on-chip models

Morrison,

Sjoerds,

Vonk

et al. 2024

Front. Immunol.

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Impressive advances have been made to replicate human physiology in vitro over the last few years due to the growth of the organ-on-chip (OoC) field in both industrial and academic settings. OoCs are a type of microphysiological system (MPS) that imitates functional and dynamic aspects of native human organ biology on a microfluidic device. Organoids and organotypic models, ranging in their complexity from simple single-cell to complex multi-cell type constructs, are being incorporated into OoC microfluidic devices to better mimic human physiology. OoC technology has now progressed to the stage at which it has received official recognition by the Food and Drug Administration (FDA) for use as an alternative to standard procedures in drug development, such as animal studies and traditional in vitro assays. However, an area that is still lagging behind is the incorporation of the immune system, which is a critical element required to investigate human health and disease. In this review, we summarise the progress made to integrate human immunology into various OoC systems, specifically focusing on models related to organ barriers and lymphoid organs. These models utilise microfluidic devices that are either commercially available or custom-made. This review explores the difference between the use of innate and adaptive immune cells and their role for modelling organ-specific diseases in OoCs. Immunocompetent multi-OoC models are also highlighted and the extent to which they recapitulate systemic physiology is discussed. Together, the aim of this review is to describe the current state of immune-OoCs, the limitations and the future perspectives needed to improve the field.

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Modeling Structural Elements and Functional Responses to Lymphatic‐Delivered Cues in a Murine Lymph Node on a Chip

Mazzaglia,

Munir,

Lei

et al. 2024

Adv Healthcare Materials

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Lymph nodes (LNs) are organs of the immune system, critical for maintenance of homeostasis and initiation of immune responses, yet there are few models that accurately recapitulate LN functions in vitro. To tackle this issue, an engineered murine LN (eLN) was developed, replicating key cellular components of the mouse LN; incorporating primary murine lymphocytes, fibroblastic reticular cells (FRCs), and lymphatic endothelial cells (LECs). T and B cells compartments are incorporated within the eLN that mimic LN cortex and paracortex architectures. When challenged, the eLN elicits both robust inflammatory responses and antigen‐specific immune activation, showing that the system can differentiate between non‐specific and antigen‐specific responses and can be monitored in real‐time. Beyond immune responses, this model also enables interrogation of changes in stromal cells, thus permitting investigations of all LN cellular components in homeostasis and different disease settings, such as cancer. Here, we present how LN behavior can be influenced by murine melanoma‐derived factors. In conclusion, the eLN model presents a promising platform for in vitro study of LN biology that will enhance understanding of stromal and immune responses in the murine LN, and in doing so will enable development of novel therapeutic strategies to improve LN responses in disease.This article is protected by copyright. All rights reserved

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On-chip human lymph node stromal network for evaluating dendritic cell and T-cell trafficking

Cited by 4 publications

References 40 publications

Mitigating reactive oxygen species production and increasing gel porosity improves lymphocyte motility and fibroblast spreading in photocrosslinked gelatin-thiol hydrogels

Mitigating reactive oxygen species production and increasing gel porosity improves lymphocyte motility and fibroblast spreading in photocrosslinked gelatin-thiol hydrogels

In vitro immunity: an overview of immunocompetent organ-on-chip models

Modeling Structural Elements and Functional Responses to Lymphatic‐Delivered Cues in a Murine Lymph Node on a Chip

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