CD105+ cell purification and identification of endothelial function
An adapted technique for the purification of vascular endothelial cells was utilized to gather vascular endothelial cells (CD105+ cells), independently from both liver cancer tissue and normal liver tissue. The liver cancer vascular endothelial cells (T-ECs, Tumor-Endothelial Cells) and normal hepatic vascular endothelial cells (N-ECs, Normal-Endothelial Cells) microscopically showed a goose-flint-like morphology (Fig. 1A). Based on flow cytometry, CD105 positive rates of T-ECs and N-ECs were 95.6 ± 1.4 % and 94.9 ± 2.3 %, respectively (Fig. 1B), increasing the sorting efficiency of positive cells to more than 95 % to minimize the effect of negative cells on subsequent experiments, using endothelial cell tube-forming assay and endothelial cell. The endothelial cell function of T-ECs and N-ECs was identified using endothelial cell tube formation and endothelial cell phagocytosis assays. As a result, we discovered that T-ECs and N-ECs could form tubular lumen-like structures on Matrigel gel (Fig. 1C) and phagocytosed ac-LDL before emitting red fluorescence (Fig. 1D).