New categorization of human vascular endothelial cells by pro- vs anti-proliferative phenotypes

Figure 4 Phenotype evaluation of human embryonic stem cell-derived vascular endothelial cells. A: VSMC-co-culture experiments of VECs generated from three lines of hESCs (KhES-1, khES-3, khES-5) at indicated passage number (n = 3); B: qRT-PCR of RGS5 message of khES-5-derived VECs [ES (KhES-5) dEC] at indicated passage numbers; C: Western blotting of RGS5 protein in KhES-1-derived VECs [ES (KhES-1) dEC] at passage 1 and passage 6; D: Immunostaining of RGS5 protein in ES (KhES-1) dEC at passage 1 (upper) and passage 4 (lower); E: ES (KhES-5) dECs were treated with hydrogen peroxide and RGS5 message expressions were examined by RT-PCR. Lane 1: no treatment; lane 2: 100 mol/L H2O2 treatment for 2 h; lane 3: 100 μmol/L H₂O₂ treatment for 24 h; lane 4: 200 mol/L H₂O₂ treatment for 24 h; lane 5: HUVEC; F: qRT-PCR of RGS5 message of ES (KhES-5) dECs subcultured in the presence of DMSO (gray column) or 10 mol/L astaxanthin (closed columns); G: VSMC-co-culture experiments of ES (KhES-5) dECs maintained with culture medium (open column), in the presence of DMSO (gray column) or 10 mol/L astaxanthin (closed column). VSMC: Vascular smooth muscle cell; VEC: Vascular endothelial cell; PCR: Polymerase chain reaction; DMSO: Dimethylsulfoxide; RGS5: Regulator of G-protein signaling 5; HUVEC: Human umbilical vein endothelial cell.