Project description:In embryo, few Endothelial cells from Aorta would be producing Haemogenic endothelium (HE) cells. These HE cells would then be progressing to take kind of a round shape and Undergo EHT. They then take a full round shape to make Intra-aortic haematopoietic clusters (IAHC) cells. These IAHC cells would then diassociate and turn into Hematopoitic Stem Cells (HSC) that ultimately would go to produce different lineage of hematopoitic (blood) cells. We wanted to study the heterogeneity of HE in human pluripotent stem cells.
Project description:Comparison of expression profiles in adipose derived MSCs (AD-MSCs) without or with transfection of siR-EID1 and shR-EID1 and cord blood-derived HSCs (CB-HSCs). After MSCs were transfected with sRNA-EID1, they could be converted into HSCs. The goal was to determine possible molecular mechanisms of MSC transdetermination. Two-condition experiments: AD-MSCs vs CB-HSCs, and AD-MSCs transfected with the combination of siR-EID1 and shR-EID1 vs AD-MSCs transfected with shR-EID1.
Project description:Hematopoietic stem cells (HSCs) are located in the bone marrow in a specific microenvironment referred as the hematopoietic stem cell niche, where HSCs interact with a variety of stromal cells. Though several components of the stem cell niche have been identified, the regulatory mechanisms through which such components regulate the stem cell fate are still unknown. In order to address this issue, we investigated how osteoblasts (OBs) can affect the molecular and functional phenotype of HSCs and vice versa. Our data showed that CD34+ cells cultured with OBs give rise to higher total cell numbers, produce more CFU and maintain a higher percentage of CD34+CD38- cells compared to control culture. Moreover, clonogenic assay and long-term culture results showed that OBs enhance HSC differentiation towards the mono/macrophage lineage at the expense of the granulocytic and erythroid ones. Finally, GEP analysis allowed us to identify several cytokine-receptor networks, such as WNT pathway, and transcription factors, as TWIST1 and FOXC1, that could be activated by co-culture with OBs and could be responsible for the biological effects reported above. Altogether our results indicate that OBs are able to affect both HPC maintenance and differentiation capacity by modulating mono/macrophage and erythroid commitment. We set up a co-culture system composed of human CD34+ cells in culture with human OBs. After coculture, CD34+ cells and the hematopoietic cell fraction were separated from OBs and analyzed by gene expression profiling (GEP), clonogenic assay and long-term culture.
Project description:Translational regulation is of paramount importance for proteome remodeling during stem cell differentiation both at the global and transcript-specific levels. In this study, we characterized translational remodeling during hepatogenic differentiation of induced pluripotent stem cells (iPSCs) by polysome profiling. We demonstrate that protein synthesis increases during exit from pluripotency, and is then globally repressed during later steps of hepatogenic maturation. This global downregulation of translation is accompanied by a decrease in the protein abundance of components of the translation machinery, which involves a global reduction in translational efficiency of terminal oligopyrimidine tract (TOP) mRNA encoding translation-related factors. Despite global translational repression during hepatogenic differentiation, key hepatogenic genes remain efficiently translated, and the translation of several transcripts involved in hepato-specific functions and metabolic maturation are even induced. We conclude that, during hepatogenic differentiation, a global decrease in protein synthesis is accompanied by a specific translational rewiring of hepato-specific transcripts.
Project description:PV-patient specific and control healthy individual-derived iPSCs were upon differentiation for 9 days into CD34+ hematopoietic progenitors treated with IFNγ, TNFα and TGFβ1 or untreated and used for transcriptome analysis. This dataset shows presence of cell-autonomous and strong non-cell autonomous JAK2V617F-dependet inflammatory footprint at the transcriptome level.
Project description:To test our hypothesis that human embryonic stem cell-derived endothelial cells(hESC-ECs) represent their in vivo counterparts, we performed transcriptional profiling experiments in which we compared the gene expression profiles of d6 CD34+KDR+ cells, d12-14 hESC-ECs and human umbilical vein endothelial cells(HUVECs). Significantly upregulated genes were identified using significance analysis of microarrays (SAM), by comparing each of these populations to undifferentiated hESCs. Differentially regulated genes from each of these populations were examined using overlap analysis;to identify which genes are uniquely expressed in each cell type and which are expressed in common, followed by functional annotation clustering of the conserved core group of genes.
Project description:Here we derive human and chimpanzee cranial neural crest cells (CNCCs) and profile histone modifications, transcription factors, chromatin accessibility and gene expression to systematically and quantitatively annotate evolutionary divergence of craniofacial cis-regulatory landscapes. Histone modifications (H3K27ac, H3K4me1, H3K4me3, H3K27me3), chromatin modifiers (p300), transcription factors (NR2F1, TFAP2A), chromatin accessibility (ATACseq) and gene expression (RNAseq) were assayed in CNCCs derived from iPSCs/ESCs from 2 chimpanzee and 3 human individuals.
Project description:To gain insights into which pathways might be dysregulated in JMML, we compared the transcriptome profiles among hiPSC and sorted CD33+ myeloid cells from control and patient samples. NS/JMML-derived CD33+ myeloid cells showed dysregulation in major biological processes. Moreover, a consistent expression pattern among sets of genes related to pluripotency and myeloid regulation was observed in the control, NS and NS/JMML CD33+ myeloid cells further supporting generally successful myelopoiesis. Total RNA obtained from hiPSC and sorted CD33+ myeloid cells (at day 14 of differentiation) from control and patient samples
Project description:The variation among induced pluripotent stem cells (iPSCs) in their differentiation capacity to specific lineages is frequently attributed to somatic memory. In this study, we compared hematopoietic differentiation capacity of 35 human iPSC lines derived from four different tissues and four embryonic stem cell lines. The analysis revealed that hematopoietic commitment capacity (PSCs to hematopoietic precursors) is correlated with the expression level of the IGF2 gene independent of the iPSC origins. In contrast, maturation capacity (hematopoietic precursors to mature blood) is affected by iPSC origin; blood-derived iPSCs showed the highest capacity. However, some fibroblast-derived iPSCs showed higher capacity than blood-derived clones. Tracking of DNA methylation changes during reprogramming reveals that maturation capacity is highly associated with aberrant DNA methylation acquired during reprogramming, rather than the types of iPSC origins. These data demonstrated that variations in the hematopoietic differentiation capacity of iPSCs are not attributable to somatic memories of their origins. Human iPSCs after hematopoietic differentiation (n = 2), human iPSCs after neural differentiation (n = 1), human iPSCs with different culture conditions (n = 8), and human iPSC line forced to express IGF2 gene (n = 1), and its control (n = 1).
Project description:This SuperSeries is composed of the following subset Series: GSE12228: Gene expression profiling among human embryonic stem cells, differerentiated EBs and adult cells GSE12229: microRNA expression profiling among human embryonic stem cells, differerentiated EBs and adult cells Refer to individual Series