Vascular endothelial growth factor (vegf)

An enzyme-linked immunosorbent assay was used to quantify the levels of VEGF and BMP-2 (Bio-Techne, MN, USA) released by the alginates. The alginate samples were depolymerized with 1 ml of citrate buffer (150 mM sodium chloride, 55 mM sodium citrate, and 20 mM EDTA in H₂O) for 15 min at 37°C. The cell culture media and depolymerized alginate samples were analyzed at the specific time points detailed above. Assays were carried out as per the manufacturer’s protocol and analyzed on a microplate reader at a wavelength of 450 nm.

Three of the five DC pig veins and three DC human veins were reconditioned.
One of the human veins was split and reconditioned with blood from
two different donors. Before the reconditioning of the ECM, frozen
DC veins were thawed at room temperature or 4 °C overnight. Human
vein femoralis were examined for valve direction to decide on the
flow in blood reactors. All veins were connected to ligands before
the reconditioning started.
Reconditioning of veins with whole
blood has previously been described by Håkansson et al.^{18 (link)} Briefly, 30–50 mL of whole blood from
human or pig donors were collected in heparin vacutainers (BD). The
blood was mixed with STEEN solution (XVIVO Perfusion) 1:1 and 0.5%
AA (Thermo Fisher). The vascular endothelial growth factor (VEGF,
80 ng/mL, Bio-Techne) and fibroblast growth factor 2 (FGF2, 10 ng/mL,
CellGenix) were added to the solution together with 5 μg/mL
acetylsalicylic acid (Sigma-Aldrich) to prevent thrombosis. Glucose
levels were monitored and adjusted to 3–8 mM during the entire
perfusion process (Figure 7). After 7 days, veins were washed, and samples were sent
for proteomics analysis. The blood used for reconditioning was species-specific
to the vein segment.