Roles of CDX2 and EOMES in human induced trophoblast progenitor cells

Abstract

Abnormal trophoblast lineage proliferation and differentiation in early pregnancy have been associated with the pathogenesis of placenta diseases of pregnancy. However, there is still a gap in understanding the molecular mechanisms of early placental development due to the limited primary trophoblast cultures and fidelity of immortalized trophoblast lines. Trophoblasts stem (TS) cells, an in vitro model of trophectoderm that can differentiate into syncytiotrophoblasts and extravillous trophoblasts, can be an attractive tool for early pregnancy research. TS cells are well established in mouse but not in humans due to insufficient knowledge of which trophoblast lineage-specific transcription factors are involved in human trophectoderm (TE) proliferation and differentiation. Here, we applied induced pluripotent stem cell technique to investigate the human trophoblast lineage-specific transcription factors. We established human induced trophoblast progenitor (iTP) cells by direct reprogramming the fibroblasts with a pool of mouse trophoblast lineage-specific transcription factors consisting of CDX2, EOMES, and ELF5. The human iTP cells exhibit epithelial morphology and can be maintained in vitro for more than 2 months. Gene expression profile of these cells was tightly clustered with human trophectoderms but not with human neuron progenitor cells, mesenchymal stem cells, or endoderm cells. These cells are capable of differentiating into cells with an invasive capacity, suggesting extravillous trophoblasts. They also form multi-nucleated cells which secrete human chorionic gonadotropin and estradiol, suggesting syncytiotrophoblasts. Our results provide the evidence that transcription factors CDX2 and EOMES may play critical roles in human iTP cell generation.

The T-box gene family: emerging roles in development, stem cells and cancer

“…Additional T-box genes in stem/progenitor cell populations As discussed earlier, Eomes plays an essential role in TE survival in the pre-implantation embryo but it is also necessary for TSC derivation and self-renewal and for the induction of TSCs from human fibroblasts (Chen et al, 2013;Kidder and Palmer, 2010;Russ et al, 2000). Furthermore, during the differentiation of ESCs, Eomes can promote either endodermal fate or cardiovascular fate depending on high or low levels of Activin/Nodal signaling, respectively (Fig.…”

What Is Trophoblast? A Combination of Criteria Define Human First-Trimester Trophoblast

“…The ELF5 promoter is hypermethylated in the following cells: 2102Ep cells, hCG-secreting hESC, and BMP4-treated hESC, TCL1, SWAN-71, and HTR-8/SVneo ( Hemberger et al., 2010 , Bernardo et al., 2011 , Novakovic et al., 2011 , Sarkar et al., 2015 ). Fibroblasts reprogrammed with CDX2 , EOMES , and ELF5 have some characteristics of trophoblast (KRT7, GATA3, and HLA-G expression), but the ELF5 hypermethylation pattern is similar to that of the parental fibroblasts ( Chen et al., 2013a ). This differential methylation between hESC and trophoblast suggests that, as in mice, the epigenetic status of human ELF5 segregates the embryonic and extraembryonic lineages.…”

Cell Adhesion Minimization by a Novel Mesh Culture Method Mechanically Directs Trophoblast Differentiation and Self-Assembly Organization of Human Pluripotent Stem Cells

“…Indeed, over reliance on animal models due to existing ethical issues associated with the use of human embryos and practical limitations arising from the use of placentas that are less than 6 weeks old have impeded the study of the molecular mechanisms underlying proliferation and differentiation of human trophoblast lineage cells (TLCs). 3 Thus, focus is increasingly shifting toward the use of trophoblasts derived from pluripotent cells as models to study early embryogenesis in humans.…”

“…10 It is not clear whether this is a result of underlying differences in genomic and epigenetic signatures between Departments of 1 Mechanical Engineering and 2 Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan. 3 Center for Stem Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan. 4 Department of Microengineering, Postgraduate School of Engineering, Kyoto University, Kyoto, Japan.…”