Project description:STRT-N is a newly optimized single-cell RNA sequencing method for studies of early genome activation in mammalian preimplantation development. Here, single embryos from the oocyte, 2-cell, 4-cell, 8-cell, blastocyst, and morula stages were sampled for experiments and were sequenced using STRT-N method.
Project description:Transcriptional profiling of common marmoset embryo stages spanning zygote to late preimplantation blastocyst was performed by single-cell RNA-seq.
Project description:Despite morphological similarities, relatively little is known about conserved developmental processes in human and mouse pre-implantation embryos. Here we provide the first comprehensive single-cell RNA-sequencing comparison of human and mouse embryos from the zygotic to the blastocyst stage. We establish a robust computational pipeline allowing us to elucidate human-specific transcriptional Programs. Importantly, we validate our RNA-sequencing findings by Immunofluorescence analysis, which further reveals differences in human and mouseembryo gene expression. For example, although key trophectoderm factors Id2, Elf5, Eomes and Tcfap2c/Ap2? are exclusively localized to this lineage in the mouse, the human orthologues are either absent or expressed in alternative lineages. Importantly, we identify several genes exclusively expressed in the human pluripotent epiblast including the transcription factor KLF17 and key components of the TGF-? signaling pathway LEFTY1, LEFTY2, NODAL and ACVRL1/ALK1 whose expression is absent from the mouse inner cell mass. Conversely, we also identify genes with conserved expression dynamics including Foxa2/FOXA2, which we show for the first time is restricted to the primitive endoderm in both human and mouse embryos. Our analysis highlights significant differences in human pre-implantation development compared to mouse and provides a molecular blueprint to understand human embryogenesis. Single-Cell RNA-seq
Project description:We report the application of single cell RNAseq analysis of bone marrow cells from mouse-mouse intraspecies blastocyst complementation and mouse-rat interspecies blastocyst complementation.
Project description:A single mouse blastomere from until 8-cell embryo can generate an entire blastocyst. Whether blastomere-like extended pluripotent stem cells (EPS cells) retain a similar generative capacity remains unknown. Here, we established a 3D differentiation system that enabled the generation of blastocyst-like structures from EPS cells (EPS-blastoids) through lineage segregation and self-organization. EPS-blastoids resembled blastocysts in morphology and cell lineage allocation. EPS-blastoids formation recapitulated key morphogenetic events during preimplantation and early postimplantation development in vitro. Upon transfer, some EPS-blastoids underwent implantation, induced decidualization, and generated live, albeit disorganized, tissues in utero. Single cell and bulk RNA-sequencing analysis revealed that EPS-blastoids contained all three blastocyst cell lineages and shared transcriptional similarity with natural blastocysts. We also provide proof-of-concept that EPS-blastoids can be generated from somatic cells via cellular reprograming. EPS-blastoids provide a unique platform for studying early embryogenesis, and pave the way to generate viable synthetic embryos using cultured cells.
Project description:Primary Objective:
* To determine whether celecoxib downregulates GATA-6 expression to upregulate 15-LOX-1 expression and induce apoptosis in human rectal tumors, researchers will measure GATA-6 and 15-LOX-1 expression, 13-S-HODE levels, and apoptosis rates in normal and colorectal polyp epithelial tissues before and after 6 months of celecoxib treatment of patients with familial adenomatous polyposis (FAP).
Project description:Here, we provide fundamental insights into early human development by single-cell RNA-sequencing of human and mouse preimplantation embryos. We elucidate conserved transcriptional programs along with those that are human specific. Importantly, we validate our RNA-sequencing findings at the protein level, which further reveals differences in human and mouse embryo gene expression. For example, we identify several genes exclusively expressed in the human pluripotent epiblast, including the transcription factor KLF17. Key components of the TGF-β signalling pathway, including NODAL, GDF3, TGFBR1/ALK5, LEFTY1, SMAD2, SMAD4 and TDGF1, are also enriched in the human epiblast. Intriguingly, inhibition of TGF-β signalling abrogates NANOG expression in human epiblast cells, consistent with a requirement for this pathway in pluripotency. Although the key trophectoderm factors Id2, Elf5 and Eomes are exclusively localized to this lineage in the mouse, the human orthologues are either absent or expressed in alternative lineages. Importantly, we also identify genes with conserved expression dynamics, including Foxa2/FOXA2, which we show is restricted to the primitive endoderm in both human and mouse embryos. Comparison of the human epiblast to existing embryonic stem cells (hESCs) reveals conservation of pluripotency but also additional pathways more enriched in hESCs. Our analysis highlights significant differences in human preimplantation development compared with mouse and provides a molecular blueprint to understand human embryogenesis and its relationship to stem cells.