Remodeling the step-wise mouse placental development by using totipotent blastomere-like stem cells [RNA-seq]
Ontology highlight
ABSTRACT: During development, totipotent blastomeres initiate the first cell fate decision to generate inner cell mass and trophectoderm. The trophectoderm forms the placenta mediating fetal-maternal communications, while placental deficiencies cause infertility and pregnancy disorders in humans. However, in vitro systems remodeling the step-wise placental development, particularly encompassing the pre-implantation phase, are still unavailable. Here, using mouse totipotent blastomere-like cells (TBLCs), we successfully realize inducing and long-term maintaining trophectoderm-like stem cells (TELSCs), and further generating placental trophoblast organoids. Interestingly, an intermediate morula trophectoderm-like cells (TELCs) were transiently induced from TBLCs, and quickly converted into TELSCs assembling blastocyst trophectoderm. In 3D culturing condition, TELSCs can form rosette-like structures at peri-implantation period, to eventually generate trophoblast organoids, in which trophoblast progenitors/giant cells, spongiotrophoblasts and syncytiotrophoblasts were all identified at the single-cell level. Transcriptomic and epigenomic analyses enable tracing the step-by-step transition from TBLCs to mature trophoblast lineages. Thus, this study provides a comprehensive differentiation system to understand and investigate placental development.
Project description:During development, totipotent blastomeres initiate the first cell fate decision to generate inner cell mass and trophectoderm. The trophectoderm forms the placenta mediating fetal-maternal communications, while placental deficiencies cause infertility and pregnancy disorders in humans. However, in vitro systems remodeling the step-wise placental development, particularly encompassing the pre-implantation phase, are still unavailable. Here, using mouse totipotent blastomere-like cells (TBLCs), we successfully realize inducing and long-term maintaining trophectoderm-like stem cells (TELSCs), and further generating placental trophoblast organoids. Interestingly, an intermediate morula trophectoderm-like cells (TELCs) were transiently induced from TBLCs, and quickly converted into TELSCs assembling blastocyst trophectoderm. In 3D culturing condition, TELSCs can form rosette-like structures at peri-implantation period, to eventually generate trophoblast organoids, in which trophoblast progenitors/giant cells, spongiotrophoblasts and syncytiotrophoblasts were all identified at the single-cell level. Transcriptomic and epigenomic analyses enable tracing the step-by-step transition from TBLCs to mature trophoblast lineages. Thus, this study provides a comprehensive differentiation system to understand and investigate placental development.
Project description:During development, totipotent blastomeres initiate the first cell fate decision to generate inner cell mass and trophectoderm. The trophectoderm forms the placenta mediating fetal-maternal communications, while placental deficiencies cause infertility and pregnancy disorders in humans. However, in vitro systems remodeling the step-wise placental development, particularly encompassing the pre-implantation phase, are still unavailable. Here, using mouse totipotent blastomere-like cells (TBLCs), we successfully realize inducing and long-term maintaining trophectoderm-like stem cells (TELSCs), and further generating placental trophoblast organoids. Interestingly, an intermediate morula trophectoderm-like cells (TELCs) were transiently induced from TBLCs, and quickly converted into TELSCs assembling blastocyst trophectoderm. In 3D culturing condition, TELSCs can form rosette-like structures at peri-implantation period, to eventually generate trophoblast organoids, in which trophoblast progenitors/giant cells, spongiotrophoblasts and syncytiotrophoblasts were all identified at the single-cell level. Transcriptomic and epigenomic analyses enable tracing the step-by-step transition from TBLCs to mature trophoblast lineages. Thus, this study provides a comprehensive differentiation system to understand and investigate placental development.
Project description:During development, totipotent blastomeres initiate the first cell fate decision to generate inner cell mass and trophectoderm. The trophectoderm forms the placenta mediating fetal-maternal communications, while placental deficiencies cause infertility and pregnancy disorders in humans. However, in vitro systems remodeling the step-wise placental development, particularly encompassing the pre-implantation phase, are still unavailable. Here, using mouse totipotent blastomere-like cells (TBLCs), we successfully realize inducing and long-term maintaining trophectoderm-like stem cells (TELSCs), and further generating placental trophoblast organoids. Interestingly, an intermediate morula trophectoderm-like cells (TELCs) were transiently induced from TBLCs, and quickly converted into TELSCs assembling blastocyst trophectoderm. In 3D culturing condition, TELSCs can form rosette-like structures at peri-implantation period, to eventually generate trophoblast organoids, in which trophoblast progenitors/giant cells, spongiotrophoblasts and syncytiotrophoblasts were all identified at the single-cell level. Transcriptomic and epigenomic analyses enable tracing the step-by-step transition from TBLCs to mature trophoblast lineages. Thus, this study provides a comprehensive differentiation system to understand and investigate placental development.
Project description:During development, totipotent blastomeres initiate the first cell fate decision to generate inner cell mass and trophectoderm. The trophectoderm forms the placenta mediating fetal-maternal communications, while placental deficiencies cause infertility and pregnancy disorders in humans. However, in vitro systems remodeling the step-wise placental development, particularly encompassing the pre-implantation phase, are still unavailable. Here, using mouse totipotent blastomere-like cells (TBLCs), we successfully realize inducing and long-term maintaining trophectoderm-like stem cells (TELSCs), and further generating placental trophoblast organoids. Interestingly, an intermediate morula trophectoderm-like cells (TELCs) were transiently induced from TBLCs, and quickly converted into TELSCs assembling blastocyst trophectoderm. In 3D culturing condition, TELSCs can form rosette-like structures at peri-implantation period, to eventually generate trophoblast organoids, in which trophoblast progenitors/giant cells, spongiotrophoblasts and syncytiotrophoblasts were all identified at the single-cell level. Transcriptomic and epigenomic analyses enable tracing the step-by-step transition from TBLCs to mature trophoblast lineages. Thus, this study provides a comprehensive differentiation system to understand and investigate placental development.
Project description:5 days after fertilisation the human embryo forms a blastocyst, comprising an outer layer of trophectoderm (TE), that gives rise to placental trophoblast cells, and the inner cell mass (ICM) which later segregates into epiblast (EPI) and primitive endoderm (PE). After implantation TE differentiates into cytotrophoblast cells (CTBs) which give rise to the multinucleated syncytiotrophoblast (STB) and invasive extravillous trophoblast cells (EVTs). A conserved molecular cascade regulates TE initiation (Gerri et al. Nature 2020), but subsequent TE development is poorly defined. To study this in greater detail we performed RNA-seq analysis of human blastocysts cultured to different stages of pre-implantation TE development: Day 5: TE appearance, Day 6: TE expansion and Day 7: TE hatching.
Project description:Trophectoderm-specific expression of Angiomotin (AMOT) in pre-implantation embryos followed by its unique expression in the post-implantation ectoplacental cone that harbors the trophoblast stem cell niche prompted our investigation on the function of AMOT in trophoblast cells. Using the in vitro trophoblast stem cell culture model, we established differentiation dependent up-regulation of AMOT expression in trophoblast cells. To understand the function of AMOT in trophoblast cells mass spectrometry-based proteomic analysis was employed to identify the AMOT interactome within the trophoblast proteome. This approach utilized immunoprecipitation of endogenous AMOT followed by fractionation on SDS-PAGE and subsequently subjecting the tryptic digested excised gel bands to mass spectrometry.
Project description:The embryo instructs the allocation of cell states to spatially regulate growth and functions. In the blastocyst, the formation and divergence of the trophoblast lineage ensures successful implantation and placental development. Here, we defined an optimal set of molecules secreted by the inner epiblast cells (inducers) that captures stable, highly self-renewing, pre-implantation-like mouse trophectoderm stem cells (TESCs). When exposed to suboptimal inducers, these stem cells form interconvertible subpopulations with reduced self-renewal, known as trophoblast stem cells (TSCs), and resembling peri-implantation progenitors. TECSs have an enhanced capacity to form blastoids that implant more efficiently in utero. We find that this enhanced capacity is due to epiblast inducers not only controlling trophoblast growth but also trophoblast secretion of WNT6/7B that stimulates uterine decidualization. Thus, the embryo leverages epiblast inductions to drive both the growth and decidualization potential of trophoblasts required for implantation and development.
Project description:The embryo instructs the allocation of cell states to spatially regulate growth and functions. In the blastocyst, the formation and divergence of the trophoblast lineage ensures successful implantation and placental development. Here, we defined an optimal set of molecules secreted by the inner epiblast cells (inducers) that captures stable, highly self-renewing, pre-implantation-like mouse trophectoderm stem cells (TESCs). When exposed to suboptimal inducers, these stem cells form interconvertible subpopulations with reduced self-renewal, known as trophoblast stem cells (TSCs), and resembling peri-implantation progenitors. TECSs have an enhanced capacity to form blastoids that implant more efficiently in utero. We find that this enhanced capacity is due to epiblast inducers not only controlling trophoblast growth but also trophoblast secretion of WNT6/7B that stimulates uterine decidualization. Thus, the embryo leverages epiblast inductions to drive both the growth and decidualization potential of trophoblasts required for implantation and development.
Project description:Blastoids, a structure similar to blastocysts in morphological and molecular level, can be applied to regeneration research. Using totipotent cells to construct blastoids will extend the information of early development to an earlier stage, and explore clues of regeneration. Totipotent blastomere-like cells (TBLCs) are a novel type of stably cultured mouse totipotent cell line generated by inhibiting spliceosomes. Here, we constructed blastoids (TBL-blastoids) in a new three-dimensional culture system using TBLCs. Morphological and transcriptomic analysis revealed TBL-blastoids contained typical morphology and key cell lineages of blastocysts and had higher degree of consistency in developmental rate and morphology compared to other blastoids. Moreover, TBL-blastoids implanted into uterus, induced decidua and even developed to embryonic tissues, indicating their in vivo developmental potential. The expansion and structures of TBL-blastoids in the IVC system also showed their in vitro developmental potential. The efficiency of generating TBL-blastoids and implantation rate suggest the necessity of TE-like component formation. Meanwhile, TBLCs can differentiate into extraembryonic cell lines directly, which provides an alternative strategy for evaluating totipotency. Furthermore, we explored the impacts of senescence, a central role in regeneration, on TBLCs and found that cellular senescence impaired the totipotency of TBLCs and the efficiency of generating blastoids. Also, the in vivo and in vitro developmental potential of TBL-blastoids were declined. In conclusion, the induction of TBLCs into blastoids and extraembryonic cells is valuable for promoting regeneration, early embryonic development study and evaluating totipotency.
Project description:Blastoids, a structure similar to blastocysts in morphological and molecular level, can be applied to regeneration research. Using totipotent cells to construct blastoids will extend the information of early development to an earlier stage, and explore clues of regeneration. Totipotent blastomere-like cells (TBLCs) are a novel type of stably cultured mouse totipotent cell line generated by inhibiting spliceosomes. Here, we constructed blastoids (TBL-blastoids) in a new three-dimensional culture system using TBLCs. Morphological and transcriptomic analysis revealed TBL-blastoids contained typical morphology and key cell lineages of blastocysts and had higher degree of consistency in developmental rate and morphology compared to other blastoids. Moreover, TBL-blastoids implanted into uterus, induced decidua and even developed to embryonic tissues, indicating their in vivo developmental potential. The expansion and structures of TBL-blastoids in the IVC system also showed their in vitro developmental potential. The efficiency of generating TBL-blastoids and implantation rate suggest the necessity of TE-like component formation. Meanwhile, TBLCs can differentiate into extraembryonic cell lines directly, which provides an alternative strategy for evaluating totipotency. Furthermore, we explored the impacts of senescence, a central role in regeneration, on TBLCs and found that cellular senescence impaired the totipotency of TBLCs and the efficiency of generating blastoids. Also, the in vivo and in vitro developmental potential of TBL-blastoids were declined. In conclusion, the induction of TBLCs into blastoids and extraembryonic cells is valuable for promoting regeneration, early embryonic development study and evaluating totipotency.