Fig. 1: Derivation of TSCs from naïve and primed hPSCs.

A Schematic representation of protocols for deriving four types of TSCs. nTSC: directly derived from naïve hPSCs; nbTSC: derived from attached naïve hPSC-derived blastoids; ccTSC: generated from primed hPSCs via chemical conversion with HDACi VPA, PD0325901, and LIF, involving an intermediate state of epigenetic reprogramming; pdTSC: induced from primed hPSCs with initial BMP4 and WNTi (IWP2) treatment to prime cells toward the trophectoderm phase. Part of the graph was created using BioRender https://biorender.com/pfs10ip. B Immunostaining of GATA3 and KRT7 in nTSCs, nbTSCs, ccTSCs, and pdTSCs. Phalloidin and DAPI staining for cytoskeleton and nucleus. Scale bar, 100 μm. C Quantitative gene expression analysis of pluripotency, syncytiotrophoblast, trophoblast, and amnion markers in the four TSC types and their parental hPSCs (primed and naïve). Fold changes are normalized to primed hPSCs. Data are presented as mean ± SD (n = 3 independent biological replicates). Significance was analyzed by two-sided Student’s t-test. *P < 0.05 compared to primed hPSCs. Source data are provided as a Source Data file. D Proliferation ratios of nTSCs, nbTSCs, ccTSCs, and pdTSCs over a 48-h interval at passage 13. Data are presented as mean ± SD (n = 3 independent biological cell populations). One-way ANOVA was used to comparing all TSCs, and no significant differences were observed. All TSCs proliferated well, with ccTSC demonstrating slightly slower proliferation rate compared to the other TSCs, though not statistically significant. Source data are provided as a Source Data file.