Journal: Biofabrication

Article Title: A biopsy-sized 3D skin model with a perifollicular vascular plexus enables studying immune cell trafficking in the skin

doi: 10.1088/1758-5090/ad5d1a

Figure Lengend Snippet: Characterization of the vascular plexus in 3D-SoC. (A) The COMSOL model displays the calculated range of shear stresses in the vascular pattern. The design recapitulates the physiological range of shear stress levels found in the cutaneous capillaries, venules, and arterioles. We divide the vasculature into three shear rate zones as low-shear (vertical interconnecting channels), mid-shear (two outermost channels, top and bottom) and high-shear (two innermost, horizontal channels). (B) Imaging of the vascular network seeded with GFP-HDBECs confirms uniform coverage of the microchannel walls. Scale bar: 1 mm; (C) Immunofluorescent staining of primary HDBECs in 3D-SoC with VE-cadherin (VECAD; white). Scale bar: 5 µ m; (D) confocal imaging of the 3D-SoC seeded with HDBECs perfused with both 20 kDa and 40 kDa dextran at time zero and sixty minutes allowing for comparison of the permeability characteristics. Scale bar: 2 mm; (E) the graph shows increased leakage of dextran in the model without HDBECs (acellular control) for both molecular weights. (F) Time-lapse transport data integrated into a COMSOL model enabled the estimation of the average permeability of the vasculature. The permeability values were determined to be 0.62 µ m s −1 for 20 kDa and 0.41 µ m s −1 for 40 kDa respectively (** = p < 0.01).

Article Snippet: Human dermal blood endothelial cells (HDBECs) (PromoCell #C-12211) and GFP-tagged HDBECs (Angio-Proteomie #cAP-0005GFP-PM) were cultured up to passage 3 in Microvascular EC Growth Medium (PromoCell #C-22020).

Techniques: Shear, Imaging, Staining, Comparison, Permeability, Control

Journal: Biofabrication

Article Title: A biopsy-sized 3D skin model with a perifollicular vascular plexus enables studying immune cell trafficking in the skin

doi: 10.1088/1758-5090/ad5d1a

Figure Lengend Snippet: Incorporation and real-time monitoring of circulating T cells in 3D-SoC. (A) Schematic representation of the stages of T cell infiltration into human skin. (B) Live immunofluorescent images showing the naïve T cells labelled with CellTracker (red) on HDBECs in the first 1–2 min (left panel; the round morphology resembles the tethering/rolling stage); between 2–5 min (middle panel; the spread morphology resembles the firm adhesion stage); and between 5–15 min (right panel; the morphology and location relative to ECs resembles the diapedesis stage). The first two images show the top view, and the right-most image shows a cross-section of the 3D-SoC. Scale bars: 5 µ m; (C) High magnification image capturing a T cell (red) with its lamellipodia squeezing between two endothelial cells (green), resembling the morphology of T cells in vivo during their movement through capillary walls (namely diapedesis). Scale bar: 2 µ m; (D) characterization of Th1 cells polarized from Naïve T cells in vitro through flow cytometry showing expression of both Interferon γ and TNFα. (E) Comparison of the attachment of the T cells to the shear stress analysis for naive and Th1 cell population. (F) Total percentage of naïve T cells and Th1 cells retained after 5 and 10 mins of flow. (G) Percentage of cells retained for distinct shear zones; HS: high-shear, MS: mid-shear, LS: low-shear. (* = p < 0.05, ** = p < 0.01, *** = p < 0.005).

Article Snippet: Human dermal blood endothelial cells (HDBECs) (PromoCell #C-12211) and GFP-tagged HDBECs (Angio-Proteomie #cAP-0005GFP-PM) were cultured up to passage 3 in Microvascular EC Growth Medium (PromoCell #C-22020).

Techniques: In Vivo, In Vitro, Flow Cytometry, Expressing, Comparison, Shear