Functional analyses of hiPSCs-derived ECs (hiPSC-ECs) at day 12 of cultivation. (A) hiPSC-ECs formed tube-like structures when seeded on matrigel. Scale bars represent 200 µm. (B) Expression analysis of CD62E, ICAM-1, and VCAM-1 transcripts using qRT-PCR in unstimulated and TNF-α-stimulated hiPSC-ECs differentiated in vitronectin-coated 12-well plates. The mRNA levels were normalized to GAPDH mRNA levels, and the results are presented relative to the expression levels in hiPSCs. Results are shown as mean + SEM (n = 3). Statistical differences were determined using the paired t-test (** p < 0.01). (C) Flow cytometric analysis of CD62E-expressing hiPSC-ECs without and with 50 ng/mL of TNF-α stimulation differentiated in vitronectin-coated 12-well plates. Results are shown as mean + SEM (n = 3). Statistical differences were determined using paired t-test (** p < 0.01). (D) Representative immunocytochemistry images of hiPSC-ECs differentiated in vitronectin-coated 12-well plates without or with subsequent TNF-α stimulation and staining with a PE-labeled CD62E-specific antibody. Scale bars represent 100 µm. Samples were fixed, processed, and imaged identically. (E) Interaction of granulocytes (calcein AM: green) with unstimulated and TNF-α-stimulated hiPSC-ECs under static conditions. (F) Numbers of attached granulocytes to unstimulated and TNF-α-stimulated hiPSC-ECs under static conditions. Results are shown as mean + SEM (n = 3). Statistical differences were determined using the paired t-test (* p < 0.01; ** p < 0.01; *** p < 0.001).