Project description:Homogenous staining regions (hsr) are cytogenetic representations of a gene amplification. They are found exclusively in tumour cells. We found a hsr in the human embryonic stem cell (hESC) line H14, this is the first report of a hsr in hESCs. FISH and CGH studies showed that the hsr was derived from chromosome 17p11.2. The gene expression analysis studies were performed to identify the genes upregulated due to the hsr by comparison the the hsr-contianing H14 cells with a karyotypically normal parent H14 cell line. Keywords: karyotypically abnormal hESC vs karyotypically normal hESC.
Project description:Homogenous staining regions (hsr) are cytogenetic representations of a gene amplification. They are found exclusively in tumour cells. We found a hsr in the human embryonic stem cell (hESC) line H14, this is the first report of a hsr in hESCs. FISH and CGH studies showed that the hsr was derived from chromosome 17p11.2. The gene expression analysis studies were performed to identify the genes upregulated due to the hsr by comparison the the hsr-contianing H14 cells with a karyotypically normal parent H14 cell line. Experiment Overall Design: H14 HSR positive embryonic stem cells and H14 HSR negative cells were hybridised to Affymetrix U133 Plus 2.0 expression arrays. Replicates were also run using H14 positive and H14 negative cells from different passages.
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:The aim of this project is to differentiate human embryonic stem cells to an extra-embryonic fate, specifically the hypoblast. This is of uttermost importance given the current lack of human hypoblast stem cells.
We hypothesized that the pluripotent characteristics of the starting human embryonic stem cell population may dictate the competency for extra-embryonic cell fate specification. Based on this hypothesis and using human embryonic stem cells maintained in different naïve-like culture regimes, we have now developed conditions that allow the differentiation of human embryonic stem cells to a stable GATA6+ SOX2- population. This suggests that these cells may be putative human hypoblast stem cells. To validate this finding here we propose to perform RNA sequencing experiments of the differentiated human embryonic stem cells. By comparing their RNA expression profile to the single cell sequencing data of the human embryo that we are currently generating, we will be able to determine the identity of our GATA6+ SOX2- cells, and establish whether they represent the in vivo human hypoblast.
Project description:The aim of this project is to differentiate human embryonic stem cells to an extra-embryonic fate, specifically the hypoblast. This is of uttermost importance given the current lack of human hypoblast stem cells.
We hypothesized that the pluripotent characteristics of the starting human embryonic stem cell population may dictate the competency for extra-embryonic cell fate specification. Based on this hypothesis and using human embryonic stem cells maintained in different naïve-like culture regimes, we have now developed conditions that allow the differentiation of human embryonic stem cells to a stable GATA6+ SOX2- population. This suggests that these cells may be putative human hypoblast stem cells. To validate this finding here we propose to perform RNA sequencing experiments of the differentiated human embryonic stem cells. By comparing their RNA expression profile to the single cell sequencing data of the human embryo that we are currently generating, we will be able to determine the identity of our GATA6+ SOX2- cells, and establish whether they represent the in vivo human hypoblast.
This dataset contains all the data available for this study on 2020-04-20.
Project description:Here we perfomed the Teratoma assay for a normal human embryonic stem cell line (H9(+Dox)), a human embryonic stem cell line with a mesendodermal differentiation bias (H9Hyb), a normal human induced pluripotent stem cell line (LU07), a human induced pluripotent stem cell line with reactivated transgenes (LU07+Dox) and a human embryonal carcinoma cell line (EC) and anayzed their gene expression. The ability to form teratomas in vivo containing multiple somatic cell types is regarded as functional evidence of pluripotency for human pluripotent stem cells (hPSCs). Since the Teratoma assay is animal-dependent, laborious and only qualitative, the PluriTest and the hPSC ScoreCard assay have been developed as in vitro alternatives. Here we compared normal hPSCs, induced hPSCs (hiPSCs) with reactivated reprogramming transgenes and human embryonal carcinoma (hEC) cells in these assays. While normal hPSCs gave rise to typical teratomas, the xenografts of the hEC cells and the hiPSCs with reactivated reprogramming transgenes were largely undifferentiated and malignant. The hPSC ScoreCard assay confirmed the line-specific differentiation propensities in vitro. However, when undifferentiated cells were analysed with PluriTest only hEC cells were identified as abnormal whereas all other cell lines were indistinguishable and resembled normal hPSCs. Our results indicate that pluripotency assays are best selected on the basis of intended downstream applications.