Fig. 6: Evaluating line to line variation of the monolayer differentiation protocol using the identified cell surface markers. | Nature Communications

Fig. 6: Evaluating line to line variation of the monolayer differentiation protocol using the identified cell surface markers.

From: Identification of cell surface markers and establishment of monolayer differentiation to retinal pigment epithelial cells

Fig. 6

a Bright field pictures of H9, HS980, and HS983a embryonic pluripotent stem cell lines after 60 days of differentiation. b Illustrative dot plots showing TRA-1-60 and CD140b expression intensity measured by flow cytometry at day 0, day 30, and day 60 of differentiation in two of the three embryonic stem cell lines tested (HS980 and HS983a). c Percentage of positive cells for TRA-1-60, CD140b, CD56, CD184, and GD2 measured by flow cytometry at days 30 and 60 of differentiation using three different embryonic stem cell lines. d Gene expression analysis of RPE genes at day 30 and day 60 of differentiation. Values are normalized to GAPDH and displayed as relative to undifferentiated hESC. Bars represent means ± SEM from three independent experiments. e Functional assays demonstrating monolayer integrity measured by transepithelial resistance (TEER), and pigment epithelium-derived factor (PEDF) polarized secretion measured by ELISA. Basal values for HS983a were not detected. The TEER value for undifferentiated hESC is shown for comparison (dashed line). Bars in all bar graphs represent means ± SEM from three independent experiments. Scale bars: a = 200 μm. Source data are provided as a Source Data file.

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