Fig. 1

The culture system used in this study to culture HUVEC alone or with H₂O₂ in the presence or absence of different treatments of DBMSCs (CMDBMSC, SFDBMEC, and ICDBMSC). CMDBMSC culture system consisted of HUVEC seeded on a surface of 6-well culture plate in a complete endothelial cell growth culture medium (untreated HUVEC) (a) or with 100 μM H₂O₂ (b) or with 100 μM H₂O₂ and 25% CM obtained from unstimulated DBMSCs (c); SFDBMSC culture system consisted of DBMSCs seeded in the upper chamber while HUVEC seeded in the lower chamber of transwell membrane culture system (d); and ICDBMSC culture system consisted of DBMSCs seeded on the reverse side of the membrane of the chamber and HUVEC seeded on the upper side of the membrane (e). For SFDBMSC and ICDBMSC, 0.4-μm pore size transwell chamber membrane was used. HUVEC were incubated with different concentrations (1%, 5%, and 25% (v/v) CM diluted in complete DBMSC growth medium) of CMDBMSC and different ratios of 1:1, 5:1, and 10:1 HUVEC:DBMSC. Cells were then cultured in a complete endothelial cell growth medium with or without 100 μM H₂O₂ for 72 h at 37 °C in a cell culture incubator