Tube formation assay
The day before the experiment, Matrigel was left to thaw overnight in a
refrigerator set at 4 °C. The frozen μ-Slide plate and gun tip were also
removed from the refrigerator. Then, 10 μl/well of Matrigel gel was
transferred to the μ-Slide plate, which was then placed into a Petri
dish of suitable size. Then, the entire Petri dish was allowed to
incubate at 37 ° C for 30 minutes. Following the digestion of T-ECs and
N-ECs from the culture flasks, the cell suspension was set to (1
~ 2) × 105 cells/ml. Prior to the
addition of cell suspension to the wells (50 μl/well), the μ-Slide plate
was removed. Subsequently, 4 to 6 hours were spent incubating the plate
at 37 ° C. Thereafter, the medium was removed and the wells were washed
with buffer. Finally, the results of the experiment were examined under
a microscope.