Figure 3

Comparison different RPE purification methods for yield and purity. (a) Overview of steps involved in the five different RPE purification methods. (b) HDFa- RPE yield of the different purification methods. S (H1): RPE cells from H1 differentiation cultures purified by scatter sorting. (c) Purified HDFa-RPE from all five methods were cultured for 6 weeks and showed typical polygonal morphology. Scale bar, 20 μm. (d) Cells obtained from scatter low fraction post-sorting showed clusters of non-RPE cells, and polygonal RPE cells (white arrow). Scale bar, 50 μm. (e and f) 6 weeks old cultured H1-RPE cells obtained from (e) scatter high and (f) low fractions of H1 (ESC) differentiation cultures. Scale bar, 20 μm. (g) Purity of cells obtained by different methods assessed by PMEL17 flow cytometry after 6 weeks in culture. Statistical analysis for (b) and (g) done using one-way ANOVA, followed by Tukey’s honest significance difference (HSD) post hoc test. ns, not significant, (*) = p < 0.05; (**) = p < 0.005, (***) = p < 0.0005, (****) = p < 0.0001.