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. 2016 May 18;5(7):946–959. doi: 10.5966/sctm.2015-0282

Figure 1.

Figure 1.

Generation of embryonic origin-specific SMC populations for a three-dimensional coculture model. (A): Schematic representation of the in vitro differentiation of embryonic origin-specific SMCs. NE was differentiated from hESCs using FSB treatment for 7 days. hESCs were also differentiated to early mesoderm in FlyB for 36 hours and subsequently to LM in FB50 or PM in Fly for 3.5 days. For further differentiation into vascular SMCs, each of the three intermediate lineages was subjected to PT treatment for 6 days and then cocultured with HUVECs. (B–D): Validation of the identity of the intermediate lineages, LM, PM, and NE, using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis of lineage-specific markers (n = 3). (E): SMC marker expression in D6 PT-treated SMCs by qRT-PCR (∗, p < .05; ∗∗, p < .01; ∗∗∗, p < .001; ∗∗∗∗, p < .0001; n = 3 independent biological replicates). Abbreviations: D, day; FB50, fibroblast growth factor 2 plus bone morphogenetic protein 4; FLy, fibroblast growth factor 2 plus LY294002; FlyB, fibroblast growth factor 2 plus LY294002 plus bone morphogenetic protein 4; FSB, fibroblast growth factor 2 plus SB431542; hESC, human embryonic stem cell; HUVECs, human umbilical vein endothelial cells; LM, lateral mesoderm; mStrawb, mStrawberry; NE, neuroectoderm; PM, paraxial mesoderm; PT, platelet-derived growth factor-BB plus transforming growth factor-β; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; SMC, smooth muscle cell.