Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Derivation of Endothelial Cells From Human Embryonic Stem Cells by Directed Differentiation

doi: 10.1161/atvbaha.110.204800

Figure Lengend Snippet: Figure 1. Taqman low-density array (TLDA) analysis of mRNA levels in pluri- potent hESCs and cells subjected to directed differentiation to vascular endo- thelial cells. hESCs were either main- tained under pluripotency conditions or under feeder- and serum-free monolayer-directed differentiation to vas- cular endothelial cells over 21 days. TLDA analysis was performed on extracted mRNA at different time points. A, Markers shown: pluripotency- associated mRNAs: Nanog, Oct 4, and Sox 2, with significant reduction in expression observed with progression of differentiation. B, early mesoderm mark- ers (brachyury, NKX2.5, Mesp1, and Mixl1) vascular endothelial cells (FLT-1, KDR, CD31, VE-cadherin, and CD34). Data are given as the meanSEM. * P0.05, **P0.01, and ***P0.001 vs the time-matched pluripotent sample. Supplemental Table I provides the data for complete TLDA analysis.

Article Snippet: Cells were incubated at 4°C overnight with the following antibodies: mouse IgG monoclonal anti–human OCT4 (1:200; Santa Cruz Biotechnology, Inc, Heidelberg, Germany); mouse IgG monoclonal anti–human Nanog (1:200; Abcam plc, Cambridge, England); mouse IgG monoclonal anti–human CD31 (1:100; Dako UK Ltd, Ely, England); and goat IgG polyclonal VE-cadherin (1:100; R&D Systems).

Techniques: TLDA Assay, Expressing

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Derivation of Endothelial Cells From Human Embryonic Stem Cells by Directed Differentiation

doi: 10.1161/atvbaha.110.204800

Figure Lengend Snippet: Figure 2. Expression of endothelial and angiogenesis-related proteins. A, FACS analysis and immunocytofluorescent staining of endothelial marker proteins. Increasing positive expression of CD31 (FL1 and far panel) and VE-cadherin (FL2 and middle panel) observed in a time-dependent manner to 21-day differen- tiation, using 4,6-diamidino-2-phenylindole (DAPI) (nuclear counterstain [blue]) (scale bar indicates 20 m). Representative scans and images from 3 independent experiments. B, Mem- brane scans of angiogenic markers in differentiated hESC-ECs (days 0, 10, 14, and 21 of differentiation). Array images are from 10-minute exposure to x-ray film. Supplemental Figure III pro- vides the corresponding densometric analysis.

Article Snippet: Cells were incubated at 4°C overnight with the following antibodies: mouse IgG monoclonal anti–human OCT4 (1:200; Santa Cruz Biotechnology, Inc, Heidelberg, Germany); mouse IgG monoclonal anti–human Nanog (1:200; Abcam plc, Cambridge, England); mouse IgG monoclonal anti–human CD31 (1:100; Dako UK Ltd, Ely, England); and goat IgG polyclonal VE-cadherin (1:100; R&D Systems).

Techniques: Expressing, Staining, Marker