Modelling stromal-compe tent human lymph nodes in vitro
With the rapid progress in the global need for humane research, the immune system remains cemented as a critical factor that requires addressing and incorporation into animal-alternative models. While the LN itself acts essentially as th e control centre of human immunology, and advancements in lymphoid or gan models have shown promising developments, incorporation of the in fluential LNSCs has yet to be properly achieved. Therefore, this thes is aims to build a 3D LN model with a stromal cell backbone in both s tatic and dynamic settings to mimic physiological LN functioning. Cha pter 2 provides a comprehensive overview of all single- and multi-OoC models that have incorporated the human immune system. This explores the level of immunocompetency for each OoC model with the use of inn ate and adaptive immune cells, while also discussing the current limi tations, challenges and degrees of physiological improvements. Before diving into the development of a LN model with an FRC component, Cha pter 3 outlines our initial efforts in examining whether it was possi ble to culture human LN-isolated FRCs ex vivo, and whether these FRCs could represent characteristics of human LNSCs subsets. In Chapter 4 , we investigate the possibility of constructing a human LN model usi ng cultured FRCs to act as the foundation for DC integration. However , the LN contains a variety of immune cells, such as a plethora of ly mphocyte and myeloid cell subsets. Therefore, in order to advance the two cell FRC-DC model into one reflecting a LN environment to study immune cell functionality, Chapter 5 reports the incorporation of nat ive LN-derived immune cells into a LN model, enriched with FRCs. As m entioned, the LN is a dynamic environment characterised by immune cel l trafficking, chemotactic gradients and cell-cell interactions. Stat ic models fail to mimic these elements, which are pivotal in facilita ting efficient and potent immune responses. Therefore, the last exper imental study of this thesis, described in Chapter 6, aimed to bring the LN model into an OoC device and establish the lymphatic vasculatu re through the LN model using autologous LECs. Finally, Chapter 7 col lectively discusses the key findings of each chapter to conclude this thesis.
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DESCRIPTION: With the rapid progress in the global need for humane res earch, the immune system remains cemented as a critical factor that r equires addressing and incorporation into animal-alternative models. While the LN itself acts essentially as the control centre of human i mmunology, and advancements in lymphoid organ models have shown promi sing developments, incorporation of the influential LNSCs has yet to be properly achieved. Therefore, this thesis aims to build a 3D LN mo del with a stromal cell backbone in both static and dynamic settings to mimic physiological LN functioning. Chapter 2 provides a comprehen sive overview of all single- and multi-OoC models that have incorpora ted the human immune system. This explores the level of immunocompete ncy for each OoC model with the use of innate and adaptive immune cel ls, while also discussing the current limitations, challenges and deg rees of physiological improvements. Before diving into the developmen t of a LN model with an FRC component, Chapter 3 outlines our initial efforts in examining whether it was possible to culture human LN-iso lated FRCs ex vivo, and whether these FRCs could represent characteri stics of human LNSCs subsets. In Chapter 4, we investigate the possib ility of constructing a human LN model using cultured FRCs to act as the foundation for DC integration. However, the LN contains a variety of immune cells, such as a plethora of lymphocyte and myeloid cell s ubsets. Therefore, in order to advance the two cell FRC-DC model into one reflecting a LN environment to study immune cell functionality, Chapter 5 reports the incorporation of native LN-derived immune cells into a LN model, enriched with FRCs. As mentioned, the LN is a dynam ic environment characterised by immune cell trafficking, chemotactic gradients and cell-cell interactions. Static models fail to mimic the se elements, which are pivotal in facilitating efficient and potent i mmune responses. Therefore, the last experimental study of this thesi s, described in Chapter 6, aimed to bring the LN model into an OoC de vice and establish the lymphatic vasculature through the LN model usi ng autologous LECs. Finally, Chapter 7 collectively discusses the key findings of each chapter to conclude this thesis. More information o n the Modelling stromal-competent human lymph nodes in vitro END:VEVENT END:VCALENDAR