Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

Figure 2

Selection of cells with Pvec activity during differentiation. (a) Schematic representation of selection strategy during differentiation of genetically modified ESCs to isolate Pvec⁺ cells. (b) Optical microscope images of clone A11- and E14T-derived cells, during selection at d4 (before the addition of puromycin), d5, and d10 (selected cells). (c) Quantification of EGFP⁺ cells during clone G11 differentiation/selection at d4.5, d6, d7, and d8 by FACS analysis (representative results are shown). (d) Analysis of FACS data from three independent experiments. . (e–g) G11-derived EGFP⁺ selected cells at d7 coexpress VE-cadherin, Isl1, and GATA4. Note that EGFP was detected at low Isl1 and GATA4 expressors. (h) Expression of mesodermal, endodermal, and neuroectodermal markers during selection of clone A11 at d3, d7, and d10 by RT-PCR analysis.

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