primary human retinal microvascular ec  (Angio-Proteomie)

Journal: Frontiers in Molecular Biosciences

Article Title: Fibroblast activation in response to TGFβ1 is modulated by co-culture with endothelial cells in a vascular organ-on-chip platform

doi: 10.3389/fmolb.2023.1160851

Figure Lengend Snippet: Validation of high flow pump and EC response to FSS. (A) Photo of the pneumatically actuated PREDICT96 high flow pump lid showing 96 arrayed micro-pumps. (B) Representative images of Hoechst stain and KLF2-GFP expression in reporter EC subjected to different FSS conditions. Insets show close-up of KLF2 for select conditions. Quantification of (C) total nuclei and (D) KLF2 expression as a function of FSS. ** p < 0.005, **** p < 0.0001 for one-way ANOVA with Tukey’s post hoc test for n = 4-8 devices per condition. (E) Comparative gene expression for shear-responsive genes in retinal microvascular ECs; positive values indicate upregulation with exposure to high FSS (7 dyn/cm 2 ) relative to low FSS (0.5 dyn/cm 2 ), while negative fold change indicates downregulation. (F) Electron micrograph showing ultrastructure of retinal microvascular EC under low FSS, exemplified with cytoplasmic organelles, including rough endoplasmic reticulum (RER; #), Golgi (+), and mitochondria (white arrow). (G) Electron micrograph of EC under physiological FSS with more abundant RER and electron dense bodies consistent with lipids are observed. Gold particle labeling indicated by black arrows. Images at x7000 magnification.

Article Snippet: Primary human retinal microvascular EC were purchased from Angio-Proteomie (Boston, MA) and expanded in the manufacturer’s Endothelial Growth Medium.

Techniques: Biomarker Discovery, Staining, Expressing, Gene Expression, Shear, Labeling

Journal: Scientific Reports

Article Title: A high-throughput microfluidic bilayer co-culture platform to study endothelial-pericyte interactions

doi: 10.1038/s41598-021-90833-z

Figure Lengend Snippet: Development of the microfluidic microvascular co-culture model in the PREDICT96 platform. ( A ) Side-view schematic of the microvasculature. Capillaries are surrounded by pericytes which, under healthy conditions, help stabilize and mature the endothelium. ( B ) Cross-section schematic showing interaction of endothelial cells (ECs) and pericytes (PCs) through basement membrane. ( C ) Side-view cross-section schematic of the vascular model in the bilayer microfluidic device. ECs and PCs are cultured on either side of a microporous membrane coated with extracellular matrix, which allows interaction between the two cell types. ( D ) Top-view of the PREDICT96 plate, containing an array of 96 bilayer microfluidic devices that interfaces with a 384 well plate top. ( E ) Schematic of PREDICT96 custom pneumatic pump lid, containing 192 individual pumps that control fluid flow in each channel of the 96 bilayer devices. ( F ) A single PREDICT96 device corresponds to 4 wells of the 384 well plate with architecture allowing for culture and fluid flow in separate top and bottom channels, which overlap in the device center. ( G ) Top-view bright field image of a single device, with channel overlap area indicated as the region of interest (dotted rectangle). ( H ) Representative image of ECs stained for PECAM-1 (green) and Hoechst (blue) in channel overlap area.

Article Snippet: Primary human retinal microvascular endothelial cells (ECs) were purchased from Angio-Proteomie (Boston, MA) and expanded in the manufacturer’s Endothelial Growth Medium (EGM).

Techniques: Co-Culture Assay, Membrane, Cell Culture, Control, Staining