Fig. 1: Development of a xeno-free culturing condition for human EPS cells.

a FACS analysis of the percentages of OCT4-Tdtomato+ H1-EPS cells with indicated factor treatment after 2 passages. n = 2 biologically independent samples. None, LCDM medium. b Analysis of cell numbers of feeder-free cultured hEPS cells under the treatments of vitamin C, catalase, and their combination. VC, vitamin C. n = 3 biologically independent samples. ES1-EPS cells were used. c Analysis of cell numbers of feeder-free cultured hEPS cells in different basal culturing media. n = 4 biologically independent samples. ES1-EPS cells were used. d–f Analysis of the effects of different matrix proteins on feeder-free cultured hEPS cells. The percentages of adherent hEPS cells at 1.5 h after seeding was shown in d. Survival of dissociated hEPS cells 24 h after seeding was shown in e. Proliferation of hEPS cells at 72 h after seeding was shown in f. For e, f, index represents the cell number at a specific time point divided by the number of seeding cells. For d–f, n = 3 biologically independent samples. ES1-EPS cells were used. g Representative morphologies of xeno-free hEPS cells with different genetic backgrounds. Similar images were obtained in at least 5 independent experiments. Error bars, mean ± SD. All differences between means with P < 0.01 are indicated. **P < 0.01; ***P < 0.001; ****P < 0.0001. Statistical significance was analyzed using one-way ANOVA with Tukey multiple comparison test in b–f. Scale bar, 100 μm. Experiments in a–g were all independently repeated at least three times with similar results.