Journal: bioRxiv

Article Title: VEGF-A/C co-stimulation, without shear stress, triggers the polarization of lymphatic microvessels

doi: 10.1101/2025.08.13.670220

Figure Lengend Snippet: (A) Scheme of the silicone LµV shown in grey, and the fabrication protocol to produce a hollow tube in collagen gels represented in pink. After LEC seeding, the endothelial tissue is cultured for three days. (B) Optical micrograph of the LµV just after its fabrication and after two days of culture with or without stimulation with growth factors. (C) The LµV contour shown in red is segmented on each side of the vessel. (D) The graph shows the relative change of the contour length after 3 days of culture with or without growth factor stimulation. (E) Live imaging of a LµV shows the dynamic formation of sprouts in response to VEGF-A/C co-stimulation. Segmented sprouts are shown in red. (F) Maximum intensity projection (MIP) of confocal micrographs showing a sprout after 3 days of VEGF-A/C co-stimulation. Labels: phalloidin in gray and nucleus in cyan. Scale bars indicate 100 µm, except for the MIP, where it represents 50 µm.

Article Snippet: Human dermal lymphatic endothelial cells (HDLECLot 483Z001.2, PromoCell) were cultured in Microvascular Endothelial Cell Growth Medium-2 BulletKit (EGM-2, Lonza) in standard tissue culture incubators at 37°C, 95% humidity, and 5% CO2.

Techniques: Cell Culture, Imaging

Journal: PLoS ONE

Article Title: Toll-Like Receptor 4 Decoy, TOY, Attenuates Gram-Negative Bacterial Sepsis

doi: 10.1371/journal.pone.0007403

Figure Lengend Snippet: LEC (primary lymphatic microvascular endothelial cells derived from human adult dermis) were purchased from Cambrex Inc. (East Rutherford, NJ) and maintained in endothelial cell basal medium-2 with growth supplements (EBM-2 MV). Passage 4–6 LEC were incubated in EBM-2 MV containing 1 % FBS for 8 hr and then with 1 µg/ml of Fc, TFE, TOY3, or TOY8 for 15 min, and then the LEC were treated with LPS (500 ng/ml) for 30 min. (A) For determination of NF-κB activation, nuclear translocalization of p65 (a subunit of NF-κB) was analyzed by immunostaining (green). Nuclei were counterstained with DAPI (blue). Arrows indicate nuclear translocalization of p65. Scale bars, 100 µm. (B) Cells positive for p65 intranuclear staining (white arrows) were counted among 100 cells arbitrarily chosen in 4 different regions, and the values presented as a percentage of the total cell number. Bars represent means ± S.D. ( n = 4). *, P <0.05 versus Fc. (C) Primary cultured macrophages from mouse peritoneal cavity were pre-treated with 1.0 µg/ml of Fc, TOY3, or TOY8 for 30 min, and then were treated with LPS (100 ng/ml) for 4 hr. Culture media were sampled, and levels of TNF-α were measured. Bars represent means ± S.D. ( n = 5). *, P <0.05 versus Fc.

Article Snippet: Briefly, LEC (primary lymphatic microvascular endothelial cells derived from human adult dermis) were purchased from Cambrex Inc. (East Rutherford) and maintained in endothelial cell basal medium-2 with growth supplements (EBM-2 MV).

Techniques: Derivative Assay, Incubation, Activation Assay, Immunostaining, Staining, Cell Culture