ABSTRACT: Differentiating embryonic stem cells into cardiomyocytes is inefficient, and we discover that FGF-10 can induce embryonic stem cells differentiation into cardiomyocytes. We use microarray to gain insight into the global gene expression and elucidate the machenism that FGF-10 induces embryonic stem cells differentiation into cardiomyocytes. Two-day embryoid bodies were treated with or without 100 ng/ml FGF10 and RNA was obtained 24 hours later and hybridized by Affymetrix microarray
Project description:Differentiating embryonic stem cells into cardiomyocytes is inefficient, and we discover that FGF-10 can induce embryonic stem cells differentiation into cardiomyocytes. We use microarray to gain insight into the global gene expression and elucidate the machenism that FGF-10 induces embryonic stem cells differentiation into cardiomyocytes.
Project description:The associated files are mass spec data from size exclusion chromatographic separations of mouse embryonic stem cells with and without RNAse A treatment.
Project description:Human pluripotent stem cell-derived kidney organoids replicate embryonic nephrogenesis in a 3D-culture system. Recent advances suggest that refining the culture environment to replicate spatiotemporal cues present during embryonic organogenesis improves patterning. Here, this paradigm was applied to FGF signalling, a key regulator of embryonic nephron progenitor maintenance, nephrogenesis and ureteric branching. Both FGF8b and FGF10 signalling is sufficient to support nephrogenesis, with each having distinct effects on nephron patterning. FGF10 enhanced the initial WT1+ mesenchymal population, leading to proximally biased nephrons, while FGF8b biased toward early distal patterning, leading to the formation of cells with connecting segment identity. The addition of both FGF8b and FGF10 together had an additive effect, leading to a balance of proximal and distal patterning. This differential patterning was retained in tissue transplanted under the murine renal capsule, with FGF8b-treated organoids displaying increased distal/connecting segments. These findings highlight plasticity during organoid nephrogenesis that can be modulated by FGF signalling and identify an approach to refine nephrogenesis toward key cell types.
Project description:Analysis of epithelial type cell formation with mouse embyronic stem cells following 11 days of differentiation using the small molcule IDE2 as well as FGF2 and FGF10 growth factors. Up-regulation of selected lung epithelial genes will provide evidence as to whether these IDE2-based differentiation protocols result in a higher yield of alveolar-epithelial type cells. Total RNA was isolated following 11 days of differentiation in culture using IDE2 with or without FGF2 or FGF 10. Undifferentiated embryonic stem cells were taken as a control
Project description:The generation of sufficient numbers of mature ventricular myocytes for effective cell-based therapy is a central barrier for cardiac regenerative medicine. Here we demonstrate that induced pluripotent stem cells (iPSCs) can be derived from murine ventricular myocytes, and consistent with other reports of iPSCs derived from various somatic cell types, ventricular myocyte derived iPSCs (ViPSCs) exhibit a markedly higher propensity to differentiate into beating cardiomyocytes as compared to genetically-matched embryonic stem cells (ESCs) or iPSCs derived from tail-tip fibroblasts. Strikingly, ViPSC-derived cardiomyocytes form up to 99% ventricular myocytes suggesting that ventricular myocyte-derived iPSCs may be a viable strategy to generate specific cardiomyocyte subtypes for cell-based therapies. The enhanced ventricular myogenesis in ViPSCs is mediated via increased numbers of cardiovascular progenitors at early stages of differentiation. In order to investigate the mechanism of enhanced ventricular myogenesis from ViPSCs, we performed global gene expression and DNA methylation analysis, which revealed a distinct epigenetic signature that may be involved in specifying the ventricular myocyte fate in pluripotent stem cells. Total RNA was extracted from mouse ES cells, tail tip fibroblasts (TTFs), ventricular myocytes (VMs), TTF-derived induced pluripotent stem cells (TiPSCs) and VM-derived induced pluripotent stem cells (ViPSCs). Global gene expression profiling was performed using affymetrix mouse 430 2.0 gene arrays.
Project description:To identify the miRNA that potentially promote the maturation of Embryonic Stem Cells-Derived Cardiomyocytes, we performed miRNA assay profiling to further know the miRNA expression that differentially expressed after coculture with endothelial cells To further know the miRNA expression that differentially expressed after coculture with endothelial cells, we performed miRNA assay profiling to identify the miRNA that potentially promote the maturation of Embryonic Stem Cells-Derived Cardiomyocytes Embryonic Stem Cells-Derived Cardiomyocytes were re-sorted from coculture system with GFP promoter
Project description:Primitive neural stem cells (NSCs) could be derived from pluripotent mouse embryonic stem (ES) cells, and then differentiate into definitive-type neural stem cells which resemble NSCs obtained from the central nervous system. Hence, primitive NSCs define an early stage of neural induction and provide a model to understand the mechanism that controls initial neural commitment. In this study, we performed microarray assay to analyze the global transcriptional profiles in mouse ES cell-derived primitive and definitive NSCs and to depict the molecular changes during the multi-staged neural differentiation process. Primitive NSCs derived directly from ESCs in Lif (p-NSC_L), primitive NSCs that were sub-cultured in the presence of Lif and FGF (p-NSC_LF), as well as definitive NSCs derived from primitive NSCs in medium containing FGF and EGF, were collected for RNA extraction and hybridization on Affymetrix microarrays. Mouse ESCs and NSCs obtained from mouse embryonic brain (E11.5) were included for controls. For each cell type, we collected two biological replicate samples for microarray analysis.
Project description:Fibroblast growth factor (FGF) 10 is essential for lung morphogenesis and polymorphisms in the Fgf10 region are linked with higher susceptibility towards COPD in human patients. We found that FGF10 signaling is impaired in lung septal wall compartment from COPD patients. Mice with impaired FGF10 signaling (Fgf10+/-) were more prone to develop cigarette smoke (CS)-induced emphysema and pulmonary hypertension (PH). Furthermore, FGF10 overexpression could successfully reverse cigarette smoke-induced emphysema and PH in mice.
Project description:DM1 and DM2 biopsies from patients were compared to Normal adult individuals Keywords: 3 groups of samples 10 DM1 biopsies, 20 DM2 biopsies, and 6 Normal individuals biopsies
Project description:A simultaneous stimulation of the Activin / FGF, BMP, and WNT pathways is required for promoting most efficient mesoderm induction in human embryonic stem cells, as well as for subsequent differentiation into cardiomyocytes. To reveal the contributions of three of these signaling pathways to mesoderm formation and cardiac induction, comparative differentiation time-courses were recorded, varying the combinations of signaling factors administered to the cells during the first day of differentiation: FGF (F) + BMP (B) + WNT (W) treatment during the first 24 hours, or FGF + BMP, or BMP + WNT, or FGF + WNT