Numerous cell types, including neurons, iPSCs (stem cells), HUVECs
(vascular endothelial cells), primary human immune cells, and tumor cells have been successfully cultured and co-cultured on Prellis' scaffolds.
Next, we further investigated whether miR-19b of EMPs could affect HUVEC
migration and angiogenesis.
The medium collected from transfected cells was used as a source of Sema3 proteins for examining effects of mutant Sema3C (R745A), with the comparison to the wild-type Sema3C, on microcapillary formation by HUVEC
cells in in vitro angiogenesis assays.
After U2OS or HUVEC
were infected in the 6-well co-culture system (0.4-^im aperture) for 48 h, HUVEC
were seeded into the lower chamber with 100,000/well and U2OS/EGFL7i into the upper chamber with the same density, followed by co-culturing for 48 h.
The viability of HUVEC
was assessed by flow cytometry with the determination of the number of viable cells, as well as those in early and late apoptosis and necrosis evaluated by Annexin-V/PI (BioLegend, San Diego, CA, USA) double staining.
In addition, we also evaluated the expression of MTA1 in two normal cell lines, HUVEC
and Het1A cells, using another MTA1 antibody (band in HUVECs
can be visualized when the dilution of the MTA1 antibody increased to 1: 500, cat: 5646, CST) and found significantly lower expression of MTA1 in both cell lines compared with MCF-7 cells (Figure S1).
Next, in in vitro HUVEC
culture, we first examined the effects of different concentrations of iodixanol and iohexol on HUVEC
proliferation/cytotoxicity after stimulation for 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 12 h, and 24 h (Figure 2(a)).
Caption: Figure 4: NOX expression in HUVEC
suppressed by anti-RAGE antibody and NOX inhibitor.
After being incubated with elaidic acid for 48 h, HUVEC
were harvested by centrifugation at 4[degrees]C (1000 xg, 5 min).
During replicative senescence of human endothelial cells, this miRNA is continuously decreased in higher passage cells and its overexpression was found to delay the appearance of the senescence-like phenotype through direct targeting of NADPH oxidase 4 (NOX4) protein, a major ROS generator in HUVEC
cells [104, 105].
However, whereas endothelial tight junction is known as an important regulator in the paracellular permeability of endothelial cells, the relevant mechanism of HUVEC
damaged by ox-LDL remains to be fully understood.
CaD significantly inhibited the proliferation of human umbilical vein endothelial cells (HUVEC
) induced by VEGF (10ng/ml), without significantly affecting HUVEC
proliferation in the absence of VEGF and reduced also VEGF-induced angiogenesis in vivo .