Project description:Human embryonic stem cells (ESCs) were differentiated into GATA3-expressing extraembryonic cells (GATA3+ ExE cells) by exposure to chemical inhibitors targeting ACTIVIN/NODAL (A83-01, 1uM) and FGF signaling (PD03714, 100nM). Using these GATA3-expressing cells as a starting material, we established trophoblast stem-like cells (TSLCs) following the established protocol for deriving trophoblast stem cells (TSCs) from the villous trophoblast of human first-trimester placenta. To characterize the resulting ESC-derived TSLCs, RNA sequencing was conducted, allowing comparisons with the parental ESCs and previously published TSCs.
Project description:In order to investigate the characteristics and mechanisms of embryonic stem cell derived exosomes attenuates transverse aortic constriction induced ventricular remodeling, the proteomic profiles of human embryonic stem cell derived exosomes were analysed by label-free quantification.
Project description:Human embryonic stem cells (ESCs) were differentiated into GATA3-expressing extraembryonic cells (GATA3+ ExE cells) by exposure to chemical inhibitors targeting ACTIVIN/NODAL (A83-01, 1uM) and FGF signaling (PD03714, 100nM). Using these GATA3-expressing cells as a starting material, we established trophoblast stem-like cells (TSLCs) following the established protocol for deriving trophoblast stem cells (TSCs) from the villous trophoblast of human first-trimester placenta. To understand the transcriptional properties of individual TSLCs, we obtained the gene expression profile from single cells.
Project description:Embryonic stem (ES) cells and embryos reversibly pause via chemical mTOR inhibition. In this study, we investigate the tissue-specific response to mTORi-induced pausing in ES and trophoblast stem (TS) cells. To resolve the sequential rewiring of the proteome, we conducted a time-series proteomics experiment at 1, 3, 6, 12, 24, and 48 hours upon induction of pausing, and at 1, 3, 6, 12, 24, and 48 hours upon release of pausing in ES and TS cells. We find that ES, but not TS cells pause reversibly. To optimise developmental pausing conditions, we reasoned that by understanding the difference in pausing response of ES and TS cells, we could identify which pathways are essential for pausing. We found that KEGG pathways related to amino acid degradation, fatty acid degradation, and DNA repair are upregulated in ES cells, but downregulated in TS cells during entry into pausing. Moreover, by targeted metabolomics, we found a depletion of short chain carnitines in the paused ES cells. To extend the length of developmental pausing, we supplemented paused embryos with L-carnitine. The L-carnitine supplementation facilitates lipid usage and prolongs the pausing length by 19 days through the establishment of a more dormant state.
Project description:Heart deformity is the leading cause of mortality in Turner syndrome (TS) patients. To investigate the dysregulated ceRNA network in cardiomyocytes (CMs) of TS patients, we employed a ceRNA microarray to profile the mRNA-lncRNA-circRNA network in induced pluripotent stem cells (iPSCs) and their derived CMs from healthy donors (WT) and TS patients.
Project description:The differentiation of pluripotent stem cells to hepatocyte-like cells offers the perspective of unlimited supply of human hepatocytes. However, the degree of differentiation of hepatic-like cells (HLC) remains controversial. To obtain an unbiased characterization, we performed a transcriptomics study using the Affymetrix Gene Chip® HG-U133 plus 2.0, with HLC derived from embryonic and induced stem cells (ESC, hiPSC) from two different laboratories. Genome-wide gene expression profiles of ESC and HLC were compared to freshly isolated and up to 14 days cultivated primary human hepatocytes (see submission E-MTAB-3994) obtained under patient informed consent from three male donors. Three biological replicas were used for each ESC and HLC models. (associated data set: E-MTAB-3994 - Gene expression profile of human embryonic stem cell-derived hepatocyte-like cells in matrigel and recombinant laminins)
Project description:Embryonic stem (ES) cells and trophoblast stem (TS) cells are both derived from early embryos, yet these cells have distinct differentiation properties. ES cells can differentiate into all three germ layer cell types, whereas TS cells can only differentiate into placental cells. It has not been determined whether TS cells can be converted into ES-like pluripotent stem (PS) cells. Here we report that overexpression of a single transcription factor, Oct4, in TS cells is sufficient to convert TS cells into a pluripotent state. These Oct4 induced pluripotent stem (OiPS) cells have the epigenetic characteristics of ES cells, including X chromosome reactivation and elevated H3K27 me3 modifications. The gene expression profile of OiPS cells and ES cells was very similar. Moreover, OiPS cells can differentiate into the three germ layer cell types in vitro and in vivo. More importantly, chimeric mice with germline transmission could be efficiently produced from OiPS cells. To our knowledge, this is the first evidence showing that only one single transcription factor could convert the non-embryonic TS cells into pluripotent stem cells with pluripotency. Gene expression profile of iPS cells and trophoblast stem cells were generated by Affymetrix Mouse Gene 1.0 ST Array. The Gene expression profile of ES cell R1 in GSE17004 was used as control. Three biological repeats were included for each line.
Project description:Embryonic stem (ES) cells and trophoblast stem (TS) cells are both derived from early embryos, yet these cells have distinct differentiation properties. ES cells can differentiate into all three germ layer cell types, whereas TS cells can only differentiate into placental cells. It has not been determined whether TS cells can be converted into ES-like pluripotent stem (PS) cells. Here we report that overexpression of a single transcription factor, Oct4, in TS cells is sufficient to convert TS cells into a pluripotent state. These Oct4 induced pluripotent stem (OiPS) cells have the epigenetic characteristics of ES cells, including X chromosome reactivation and elevated H3K27 me3 modifications. The gene expression profile of OiPS cells and ES cells was very similar. Moreover, OiPS cells can differentiate into the three germ layer cell types in vitro and in vivo. More importantly, chimeric mice with germline transmission could be efficiently produced from OiPS cells. To our knowledge, this is the first evidence showing that only one single transcription factor could convert the non-embryonic TS cells into pluripotent stem cells with pluripotency.