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. 2023 Jul 3;12(13):1765. doi: 10.3390/cells12131765

Figure 2.

Figure 2

Flux-conditioned E8 (FC-E8) medium maintains pluripotent hiPSC and hESC lines. (a) Timeline of the cell culture and the application of FC-E8 medium and mixed media. (b) Brightfield pictures of the hiPSC lines and hESC line cultured for 24 h in commercial E8 medium (Gibco), in FC-E8 medium, and in mixed media (50% and 75% FC-E8). Scale bar: 500 µm. A magnified version of these pictures is provided as Supplementary Figure S5. (c) Gene expression of pluripotency markers (Nanog, TERT, and CDH2) in two hiPSC lines and a hESC line cultured for 24 h in FC-E8 medium (50%, 75%, or 100%), expressed as fold change compared to the control condition in commercial E8 medium (Gibco, Life Technologies). The data are normalized on the housekeeping gene ACTG1. A fold change of 1 indicates equal gene expression between the sample and the control condition (cell cultured in commercial E8). (d) Immunofluorescent staining of pluripotency markers (SSEA-4, OCT-3/4) in hiPSC lines and hESC line cultured for 4 days in commercial E8 medium and in FC-E8 medium. Scale bar: 50 µm. A magnified version of these pictures is provided as Supplementary Figure S7. (e) Results of the unlabeled metabolomics analysis on hiPSC lines and hESC line cultured for 24 h in commercial E8 medium, FC-E8 medium, and mixed media (50% FC-E8, 75% FC-E8) (n = 4).