Generation of human induced trophoblast stem cells

Abstract

SUMMARYHuman trophoblast stem cells (hTSC) derived from blastocysts and first-trimester cytotrophoblasts offer an unprecedented opportunity to study the human placenta. However, access to human embryos and first trimester placentas is limited thus preventing the establishment of hTSC from a variety of genetic backgrounds associated with placental disorders. In the present study, we show that hTSC can be generated from numerous genetic backgrounds using post-natal cells via two alternative methods: (I) somatic cell reprogramming of adult fibroblasts using the Yamanaka factors, and (II) cell fate conversion of naive and extended pluripotent stem cells. The resulted induced and converted hTSC (hiTSC/hcTSC) recapitulated hallmarks of hTSC including long-term self-renewal, expression of specific transcription factors, transcriptome-side signature, and the potential to differentiate into syncytiotrophoblast and extravillous trophoblast cells. We also clarified the developmental stage of hTSC and show that these cells resemble post-implantation NR2F2+ cytotrophoblasts (day 8-10). Altogether, hTSC lines of diverse genetics origins open the possibility to model both placental development and diseases in a dish.HighlightsReprogramming of human somatic cells to induced hTSC with OSKMConversion of naive and extended hPSC to hTSCGenetic diversity of hTSC linesDevelopmental matching of hTSC in the peri-implantation human embryo

Podocalyxin‐Like Protein 1 Regulates Pluripotency through the Cholesterol Biosynthesis Pathway

“…A distinguishing feature of totipotent‐like cells is the ability to differentiate into extra‐embryonic trophectoderm (TE) lineage like trophoblast stem cells (TSCs), where, for example, EPSCs have been suggested to differentiate into TSCs. [ 22 ] In our experiments, both untreated control and Dox‐treated (PODXL‐overexpressing) hEPSCs were able to differentiate into expanded cobblestone‐shape colonies reminiscent of hTSCs (CT27, derived from the early placenta; bTS11, derived from blastocysts) [ 23 ] in two passages (P2), while Dox‐treated hEPSCs exhibited higher expression of TSC markers (Figure 5D,E and Figure S5A, Supporting Information). In addition, both Dox‐treated or untreated control hEPSCs led to well‐differentiated teratomas with three embryonic germ layers and the immunohistological features of human beta‐chorionic gonadotropin (hCG) positive staining and hemorrhage areas, whereas hCG positive staining was not observed in teratomas derived from primed hESCs (Figure 5F and Figure S5B, Supporting Information).…”

“…CT27 and bTS11 are TSC‐positive controls. [ 22 ] CT27: TSCs derived from a trimester placenta, bTS11: TSCs derived from a blastocyst. E) qRT‐PCR analyses for trophoblast marker genes in primed RUES2 hESCs, RUES2‐EPSCs −Dox, and RUES2‐EPSCs +Dox cultured in hTSC medium were measured.…”

“…These results are consistent with a previous report showing that human LCDM‐EPSCs can differentiate into hTSCs akin to day 8 trophoblasts and further differentiate into TE‐like derivatives under proper induction. [ 22 ] In vivo experiments using hPSCs to generate human‐animal chimeras provide a platform to study early human embryogenesis and potentially regenerate human tissues and organs for replacement therapies. To our knowledge, we are the first to dissect the transcriptome landscape and differentiation trajectories of human–mouse chimeras at the peri‐implantation embryo stage by using scRNA‐seq.…”

An in vitro stem cell model of human epiblast and yolk sac interaction

“…This suggests that the starting hEPSC population may contain subpopulations with largely contrasting pluripotent states that span from naïve to primed. This is mirrored during human trophoblast stem cell induction wherein hEPSCs respond heterogeneously ( Castel et al, 2020 ).…”

Modeling the human placental barrier to understand Toxoplasma gondii´s vertical transmission

“…One source of fetal cell models used to study congenital transmissions are the mesenchymal stem cells isolated from human umbilical cord. During infection with T. gondii these cells are induced towards autophagic cell death by a mechanism that involves downregulation of mitochondrial stress factor Mcl-1 ( Chu et al., 2017 ).…”