Dataset Fig4e_bilaminoid_without-nTB_3D
Project
379-Okubo-bilaminoid
Title
3D images of bilaminar organoid generated from co-culture of human pluripotent stem cells (hPSC) and hPSC-derived hypoblast-like cells
Description
Recently, several studies using cultures of human embryos together with single-cell RNA-seq analyses have revealed differences between humans and mice, necessitating the study of human embryos. Despite the importance of human embryology, ethical and legal restrictions have limited post-implantation-stage studies. Thus, recent efforts have focused on developing in vitro self-organizing models using human stem cells. In this paper, the authors report genetic and non-genetic approaches to generate authentic hypoblast cells (naive hPSC-derived hypoblast-like cells (nHyCs))-known to give rise to one of the two extraembryonic tissues essential for embryonic development-from naive human pluripotent stem cells (hPSCs). Their nHyCs spontaneously assemble with naive hPSCs to form a three-dimensional bilaminar structure (bilaminoids) with a pro-amniotic-like cavity. In the presence of additional naive hPSC-derived analogues of the second extraembryonic tissue, the trophectoderm, the efficiency of bilaminoid formation increases from 20% to 40%, and the epiblast within the bilaminoids continues to develop in response to trophectoderm-secreted IL-6. Furthermore, they show that bilaminoids robustly recapitulate the patterning of the anterior-posterior axis and the formation of cells reflecting the pregastrula stage, the emergence of which can be shaped by genetically manipulating the DKK1/OTX2 hypoblast-like domain.
Funding
This work was supported by MEXT KAKENHI (grant numbers: JP16H02465, JP15H02360, JP20H05361, JP20H05762, JP21K19189, JP23K18340) and AMED (grant numbers: JP21bm0704035, JP21bm0104001, JP21gm1310011, JP23bm1323001) to Y.T.; MEXT KAKENHI (grant number: JP19H03418) and the JST FOREST Program (grant number: JPMJFR206C), JST CREST (grant number: JPMJCR2023) and AMED (grant numbers: JP17gm1110004, JP19gm1310002, JP19bm0704032, JP21bm0104001, JP21gm1310011) to T.Y.; MEXT KAKENHI (grant numbers: JP15H02360, JP19H05759) to E.S.; AMED (grant number: JP22bm1004002) to H.N.; the Takeda Science Foundation to Y.T.; the Naito Foundation to Y.T.; the Mochida Memorial Foundation for Medical and Pharmaceutical Research to Y.T.; and the Astellas Foundation for Research on Metabolic Disorders to Y.T.
Title
3D-rendered image of bilaminoid on day 6 in the absence of nTB
Description
3D-renderd image of bilaminoid generated spontaneously by the mixture of naive hPSCs (human pluripotent stem cells) and nHyCs (naive hPSC-derived hypoblast-like cells expressing GFP) on day 6. The bilaminoid was generated in the absence of nTB (trophectoderm/trophoblast-like cells). After fixation, the bilaminoid was stained for PAR6, GFP, and DAPI.
Description
Bilaminar embryo-like aggregates (bilaminoids) generated by the mixture of naive hPSCs (human pluripotent stem cells) and nHyCs (naive hPSC-derived hypoblast-like cells).
Extrinsic variables
Doxycycline was applied to the culture to induce the overexpression of GATA6 in naive hPSC-derived hypoblast-like cells
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Protein names
Partitioning defective 6 homolog alpha (
Q9NPB7)
Contrast enhancing methods
Resolution enhancing methods
Sample preparation methods