human lymph node lymphatic endothelial cells (lecs)  (ScienCell)

Journal: APL Bioengineering

Article Title: Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change

doi: 10.1063/5.0247363

Figure Lengend Snippet: Lymph node stroma model development. We sought to develop a model of immune cell egress from the lymph node (a). A representative immunofluorescence image of a portion of a normal human lymph node is shown. In human lymph nodes, lymphatics (PDPN+, CD31+, visualized in red and green) and fibroblastic reticular cells (PDPN+, in green) form the structure of the T cell zone (b). Schematic of an intended structure of the T cell zone model (c). LECs were seeded on the underside of a tissue culture inset. PhotoHA-collagen gels laden with FRCs are crosslinked above the LECs and then incubated for 30 min (d). After thermal cross-linking, the final gel consists of hyaluronic acid and collagen (e). With this methodology, FRCs formed networks (f) and LECs formed an intact monolayer (g). Scale bars are 50 μ m. Magnetic resonance imaging (MRI) demonstrates altered fluid transport in the presence of LN stroma (h), (i). Divergence of fluid was significantly decreased in the presence of LN stroma (j). Each data point represents a biological replicate (n = 3). Significance was determined by Students' t-test, with significant p values (<0.05) reported on the graph.

Article Snippet: Human lymph node lymphatic endothelial cells (LECs) (Sciencell) were cultured on fibronectin coated flasks in VascuLife® VEGF-Mv Endothelial Complete media, containing 5% fetal bovine serum and additional supplements as provided by the supplier (Lifeline Cell Technology).

Techniques: Immunofluorescence, Incubation, Magnetic Resonance Imaging

Journal: APL Bioengineering

Article Title: Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change

doi: 10.1063/5.0247363

Figure Lengend Snippet: LEC barrier integrity is decreased under IFF. Pressure heads of media are utilized to drive IFF (a). To create the highest magnitude flow rate, PE50 tubing was secured onto the tissue culture inset with no leaks (b). 3D printed lid raisers to the specifications of a 12 well plate (c) maintain sterility in the incubator. LEC proliferation is unchanged by IFF, as quantified by % of EdU+ cells (d). Representative images show LECs visualized with CD31 (gray). Nuclei are stained with DAPI (blue). Scale bars are 100 μ m (e). LEC coverage (f) and disrupted junctions (g) are quantified. Each point represents a biological replicate from an independent experiment, for n = 3. Each data point represents a biological replicate (n = 3). Significance was determined by two-way ANOVA followed by Tukey's t-test, with significant p values (<0.05) reported on each graph.

Article Snippet: Human lymph node lymphatic endothelial cells (LECs) (Sciencell) were cultured on fibronectin coated flasks in VascuLife® VEGF-Mv Endothelial Complete media, containing 5% fetal bovine serum and additional supplements as provided by the supplier (Lifeline Cell Technology).

Techniques: Sterility, Staining

Journal: APL Bioengineering

Article Title: Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change

doi: 10.1063/5.0247363

Figure Lengend Snippet: Presence of T cells disrupts LEC junctions regardless of flow and ameliorates flow-induced changes to FRCs. LECs are visualized in representative images with CD31 in gray and DAPI in blue (scale bar 100 μ m), and FRCs are visualized with F-actin in green (scale bar 50 μ m), in the presence of naïve CD8+ T cells (blue) (a). Quantification of LEC monolayer coverage (b) and disrupted junctions (c). FRC coverage in the presence of T cells and IFF (d). Scale bar is 50 μ m. Each data point represents a biological replicate (n = 3). Significance was determined by two-way ANOVA followed by Tukey's t-test, with significant p values (<0.05) reported on each graph.

Article Snippet: Human lymph node lymphatic endothelial cells (LECs) (Sciencell) were cultured on fibronectin coated flasks in VascuLife® VEGF-Mv Endothelial Complete media, containing 5% fetal bovine serum and additional supplements as provided by the supplier (Lifeline Cell Technology).

Techniques: