primary adult human dermal lymphatic endothelial cells  (PromoCell)

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Apolipoprotein A‐I Modulates Atherosclerosis Through Lymphatic Vessel‐Dependent Mechanisms in Mice

doi: 10.1161/JAHA.117.006892

Figure Lengend Snippet: Effect of lipid‐free apoA‐I treatment on platelet adhesion to lymphatic endothelial cells. A through C, Human platelets were isolated and incubated with a confluent monolayer of primary HMVEC ‐ dLyA d for 1 hour at 37°C. LEC s and platelets were identified by immunofluorescence using DAPI and anti‐ CD 61 antibodies, respectively. A, Representative images indicating that, under static conditions, apoA‐I‐treated LEC s display a “bridge effect” mediated by platelets pseudopodia, thus assembling LEC s together. B, Representative images and (C) quantification of the number of adhered platelets interacting with HMVEC in BSA ‐ (upper panel) and apoA‐I‐ (lower panel) treated LEC s. Results are the averages of 5 independent experiments. * P <0.05, as determined by 1‐tailed t test. D and E, Human platelets were isolated and perfused over primary HMVEC ‐ dLyA d seeded at maximum confluence in tissue culture treated flow chambers, at a wall shear rate of 50/s at 37°C for 8 minutes. D, LEC s and platelets were identified using DAPI and anti‐ CD 61 antibodies, respectively. E, The % augmentation in the number of adhered platelets following treatment is indicated above the bars. P =0.043, using a Wilcoxon signed rank test. Scale bars=10 μm (A and B) and 100 μm (E). apoA‐I indicates apolipoprotein A‐I; DAPI, 4′,6‐diamidino‐2‐phenylindole; HMVEC‐dLyAd, human dermal lymphatic microvascular endothelial cells‐adult; LECs, lymphatic endothelial cells.

Article Snippet: Primary human dermal lymphatic microvascular endothelial cells‐adult (HMVEC‐dLyAd) were cultured according to the manufacturer's protocol (Lonza) in EBM‐2 medium containing the EGM‐2 MV SingleQuots.

Techniques: Isolation, Incubation, Immunofluorescence, Shear

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Apolipoprotein A‐I Modulates Atherosclerosis Through Lymphatic Vessel‐Dependent Mechanisms in Mice

doi: 10.1161/JAHA.117.006892

Figure Lengend Snippet: Effect of lipid‐free apoA‐I treatment on podoplanin and VEGFR 3 expression. Primary HMVEC ‐ dLyA d were seeded at maximum confluence and incubated for 20 hours with BSA or apoA‐I. Cells were then analyzed by Western blotting with the use of (A) podoplanin and (B) VEGFR 3. Podoplanin protein was observed at 37 kDa and VEGFR 3 was observed at 170 kDa. Experiments were performed with 6 replicates per experimental group. * P <0.05, as determined by 1‐tailed t test. apoA‐I indicates apolipoprotein A‐I; HMVEC‐dLyAd, human dermal lymphatic microvascular endothelial cells‐adult; VEGFR, vascular endothelial growth factor receptor.

Article Snippet: Primary human dermal lymphatic microvascular endothelial cells‐adult (HMVEC‐dLyAd) were cultured according to the manufacturer's protocol (Lonza) in EBM‐2 medium containing the EGM‐2 MV SingleQuots.

Techniques: Expressing, Incubation, Western Blot