绒毛滋养层/trophoblast stem cell

Bipotent human trophoblast stem cells (hTSCs) recapitulate placenta development at 1st trimester. The molecular roadmap of bi-directional differentiation from hTSCs at single cell resolution remains elusive. Here, using scRNA-Seq, scATAC-Seq (assay for transposase-accessible chromatin) and Stereo-seq (SpaTial Enhanced REsolution Omics-sequencing) to determine the origin, heterogeneity, and differentiation capacity of hTSCs derived from 1st trimester villi, we firstly inferred ITGA6+/TP63+ cytotrophoblast cells (CTBs) as hTSCs. Then, time-course single-cell profiling of trophoblast cell fate decision from hTSCs enable us to comprehensively map a stepwise and multiple transcription factors involved trophoblast lineage restriction process. Furthermore, we revealed the profound influence of Ipatasertib and Ravoxertinib on hTSCs differentiation including cellular morphology, composition, and maturation through PI3K-Akt and ERK-MAPK pathway, respectively. Taken together, these data highlight a single cell-resolution roadmap for bipotent hTSCs differentiation and accelerate the development of early intervention strategy by small chemical inhibitors in placenta developmental diseases.