Fig. 1

Generation of human SSEA-1⁺ cells in vitro. a Overview of the protocol used to obtain SSEA-1⁺ and SUSD2⁺ cells. b Light microscopy images of primary endometrial cells, P3 SUSD2⁺ and SSEA-1⁺ cells cultured in TEM at day 1 and day 5. Blue arrow, EpCAM⁺ cells; red arrow, EpCAM⁻ cells. Scale bars, 100 µm. c Immunofluorescence analyses demonstrating the expression of SUSD2 and SSEA-1. Scale bars, 50 µm in 20X, 20 µm in 60X. d Light microscopy images show typical 6-well plates of clone formation of SUSD2⁺ and SSEA-1⁺ cells by crystal violet staining. Scale bars, 1 cm in 1X, 100 µm in 10X. e SUSD2⁺ cells showed higher colony-forming unit numbers than SSEA-1⁺ cells (Error bars represent s.d.; n = 3 technical replicates; two-tailed unpaired t-test, **P < 0.01). f CCK-8 analyses demonstrating higher cell proliferation of SUSD2⁺ cells. (Error bars represent s.d.; n = 3 technical replicates from one donor; two-way ANOVA, P < 0.001). g Doubling time calculated for SUSD2⁺ and SSEA-1⁺ cells. Error bars represent s.d.; n = 3 technical replicates, *P < 0.05. h Flow cytometric analysis showing the proportion of positive-cells in the populations. Red, positive-cells; blue, negative controls