ten days then the cells were placed in the 24 well cell culture plate paved with matrigel and placed at 37 u00b0 c the cells were incubated in the co2 culture box for 60 minutes the cells of each group were suspended in the dmem high glucose culture medium containing no factor according to the density of 4 5 u00d7 104 cells well and inoculated in the 24 well cell culture plate paved with matrigel the cell culture plate was placed at 37 u00b0 c in a co2 culture box for culturing for 4 to 96 hours the vascularization conditions of the blood vessel endothelial cells on the matrigel were observed under an inverted phase contract microscope respectively on the 6th hour 12th hour 24th hour and 48th hour the results showed that in the detection of in vitro vascularization the cells of the experimental group of 2 3 dihydro 3 hydroxymethyl 6 amino 1 4 benzoxazine with the concentration of 1 20 u03bcm can form varying degrees of cavity like vessel like structures on matrigel wherein the embryonic stem cells induced with 2 3 dihydro 3 hydroxymethyl 6 amino 1 4 benzoxazine with the concentration of 10 u03bcm can prominently form blood vessels on matrigel 10 u03bcm treatment group see fig4 the embryonic stem cells were evenly grown in a 24 well plate according to the density of 2 u00d7 104 cells well and cultured for 24 hours with dmem high glucose containing leukemia inhibitory factor l glutamine streptomycin u03b2 mercaptoethanol non essential amino acids nucleotides and 15 fbs the waste liquid was discarded after the cells adhered on the wall and the cells were cleaned with 1 u00d7 pbs the normal group was set the group was cultured under the culture condition that leukemia inhibitory factor was removed.