Efficient derivation of human trophoblast stem cells from primed pluripotent stem cells

Abstract

Human trophoblast stem cells (hTSCs) provide a valuable model to study placental development and function. While primary hTSCs have been derived from embryos/early placenta, and transdifferentiated hTSCs from naïve human pluripotent stem cells (hPSCs), the generation of hTSCs from primed PSCs is problematic. We report the successful generation of TSCs from primed hPSCs and show that BMP4 substantially enhances this process. TSCs derived from primed hPSCs are similar to blastocyst-derived hTSCs in terms of morphology, proliferation, differentiation potential, and gene expression. We define the chromatin accessibility dynamics and histone modifications (H3K4me3/H3K27me3) that specify hPSC-derived TSCs. Consistent with low density of H3K27me3 in primed hPSC-derived hTSCs, we show that knockout of H3K27 methyltransferases (EZH1/2) increases the efficiency of hTSC derivation from primed hPSCs. Efficient derivation of hTSCs from primed hPSCs provides a simple and powerful model to understand human trophoblast development, including the pathogenesis of trophoblast-related disorders, by generating disease-specific hTSCs.

Induction of human trophoblast stem-like cells from primed pluripotent stem cells

“…Very recently, Wei et al., ( 22 ) showed that primed PSCs convert to TSLCs under TSCM condition. They also showed that 10–12 d of BMP4 treatment could enhance the conversion process.…”

“…1 B . Although 10 to 12 d of BMP4 treatment increased the conversion, the resultant cells showed a lower efficiency of EVT differentiation based on HLA-G expression ( 22 ). Interestingly, in our hands, longer BMP4 treatment gradually increased the expression of amnion genes ( Fig.…”

“…This would determine whether the converted cells are truly amnion-like cells or not. Of note, several attempts ( 18 – 22 ) recently have been made to generate self-renewing human TSLCs from PSCs using human TSC culture medium (TSCM) ( 3 ). Recent papers reported that human PSCs can generate trophectoderm lineage stem cells under a micromesh culture condition ( 19 ) or chemically defined culture conditions ( 23 ).…”

“…Subsequent studies have reported that TSLCs can be generated from human naïve PSCs or expanded potential stem cells (EPSCs) ( 14 , 15 , 18 , 20 , 21 ). Most recently, a study showed that TSLCs can be derived from human primed PSCs under TSCM condition only, and 10 to 12 d of BMP4 treatment could enhance the conversion efficiency ( 22 ). Although it is unclear which protocol is better to generate functional TSLCs comparable to bona fide TSCs, all these reports provide valuable models to understand human trophoblast lineage development.…”

Leveraging Optimized Transcriptomic and Personalized Stem Cell Technologies to Better Understand Syncytialization Defects in Preeclampsia

“…The original TSCs described by Okae et al, however, were neither patient-specific nor could they model diseases such as EOPE that clinically manifest late in pregnancy. In response to these limitations, placental researchers, including those interested in EOPE, have begun to combine TSC and iPSC technologies ( Li et al, 2019 ; Castel et al, 2020 ; Dong et al, 2020 ; Guo et al, 2021 ; Io et al, 2021 ; Mischler et al, 2021 ; Wei et al, 2021 ) and have even begun to generate TSCs directly from human fibroblasts ( Liu et al, 2020 ). Although the methods have yet to be compared, so it remains unclear which is superior for the generation of bona fide TSCs, these methods should ultimately provide necessary pregnancy disease specific in vitro TSC models.…”