coronary artery ecs  (PromoCell)

Journal: bioRxiv

Article Title: Co-Therapy with S1P and Heparan Sulfate Derivatives to Restore Endothelial Glycocalyx and Combat Pro-Atherosclerotic Endothelial Dysfunction

doi: 10.1101/2024.11.06.622347

Figure Lengend Snippet: In vitro cellular remodeling as indicated by EC morphology via cell alignment in relation to flow. Representative phase contrast images of HCAECs at 10x are shown for all cohorts, which include HCAECs exposed to: A) static condition with no treatment, B) atheroprone region with no treatment, C) atheroprotective region with no treatment, D) static condition with co-treatment, (E) atheroprone region with co-treatment, and (F) atheroprotective region with co-treatment. All static and dynamic conditions (12 dynes/cm 2 ) were for 12 hours. Cell alignment was analyzed in two ways: (G) Object Orientation Parameter (OOP), which determines cell alignment in relation to flow and (H) Median Pairwise Alignment (MPA), which determines cell alignment in relation to other ECs. Each data point on the graph represents the mean individual OOP or MPA from one flow experiment, in which each cohort as range of n = 16 – 20. Box and whisker plot in (G) for OOP shows the minimum, first quartile, median, third quartile, and maximum values for each condition. MPA in (H) shows the mean and error bars which represents the SEM. Statistical analysis was performed using a two-way ANOVA. Significance is denoted by asterisks: *p<.05, **p<.01, ***p<.001, and ****p<.0001.

Article Snippet: Human coronary arterial ECs (HCAECs) purchased from PromoCell (Heidelberg, Germany) were cultured between passages 4 and 8 in PromoCell Endothelial Cell Growth Medium MV2 supplemented with growth factors, fetal calf serum, and antibiotic penicillin streptomycin.

Techniques: In Vitro, Whisker Assay

Journal: bioRxiv

Article Title: Co-Therapy with S1P and Heparan Sulfate Derivatives to Restore Endothelial Glycocalyx and Combat Pro-Atherosclerotic Endothelial Dysfunction

doi: 10.1101/2024.11.06.622347

Figure Lengend Snippet: Effect of co-treatment on S1PR1 (green) expression in HCAECs after 12 hours of flow at 12 dynes/cm 2 . Shown are representative 63x images of S1PR1 expression in HCAECs amongst different experimental groups. S1PR1, a G-protein coupled receptor found specifically on ECs, was used to determine its involvement in the therapeutic mechanism of action, and its expression increased in the atheroprotective and atheroprone regions when the co-treatment is introduced to the flow system, signifying activation. Blue represents cell nuclei labeled with DAPI. The scale bar is 20 μm. HCAECs in the no treatment group under A) static conditions, B) atheroprone conditions, and C) atheroprotective conditions. HCAECs in the co-treatment group under D) static conditions, E) atheroprone conditions, and F) atheroprotective conditions. G) Graph shows the mean ± SEM of percent area coverage of S1PR1 staining normalized to the control (0 hr static condition; n = 4). Statistical analysis was performed using a two-way ANOVA. Significance is denoted by asterisks: *p<.05, **p<.01, ***p<.001, and ****p<.0001.

Article Snippet: Human coronary arterial ECs (HCAECs) purchased from PromoCell (Heidelberg, Germany) were cultured between passages 4 and 8 in PromoCell Endothelial Cell Growth Medium MV2 supplemented with growth factors, fetal calf serum, and antibiotic penicillin streptomycin.

Techniques: Expressing, Activation Assay, Labeling, Staining, Control