MicroRNA profiling by array of a directed differentiation time course converting human embryonic stem cells (hES) into immature pancreatic beta cell precursors
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ABSTRACT: miRNA transcript profiling of a directed differentiation time course converting human embryonic stem cells (hES) into immature pancreatic beta cell precursors.
Project description:Transcriptional profiling of a directed differentiation time course converting human embryonic stem cells (hES) into immature pancreatic beta cell precursors.
Project description:H3K4me3 ChIP-Seq profiling of a directed differentiation time course converting human embryonic stem cells (hES) into immature pancreatic beta cell precursors. CyT49, a Viacyte proprietary male hESC line with normal karyotype was used.
Project description:Genomic microarray analysis of adrenergic-deficient (Dbh-/-) vs. wild-type control (Dbh+/+) mouse heart expression at embryonic day 10.5 (E10.5). Four Dbh-/- and four Dbh+/+ E10.5 hearts were isolated, and RNA was extracted from each for genomic microarray analysis using Affymetrix 430A 2.0 arrays. The biological samples were collected and prepared in Dr. Ebert's laboratory at the Univ of Central Florida in Orlando, FL. The RNA samples were sent to GeneLogic (Gaithersburg, MD) for the microarray analysis.
Project description:Human tissue based proteomics projects are challenging due to low abundance of proteins and tissue specificity of protein expression. In this study, we aimed to develop a cell-based approach to profile the male specific region of the Y chromosome (MSY) proteins. First, we profiled the expression of 23 Y chromosome genes and 15 of their X-linked homologues during neural cell differentiation from NT2 cells at three different developmental stages using qRT-PCR, western blotting and immunofluorescent (IF) techniques. The expression level of 12 Y-linked genes significantly increased over neural differentiation. Including RBMY1, EIF1AY, DDX3Y1, HSFY1, BPY2, PCDH11Y, UTY, RPS4Y1, USP9Y, SRY, PRY, and ZFY. Subsequently, DDX3Y was selected as a candidate for knockdown as it was significantly expressed in neural progenitor cells and it is known to be expressed in a gender specific manner and play a role in spermatogenesis. A siRNA-mediated DDX3Y knockdown in neural progenitor cells impaired cell cycle progression and increased apoptosis, consequently interrupting differentiation. Label-free quantitative shotgun proteomics based on a spectral counting approach was then used to characterize the proteomic profile of the cells after DDX3Y knockdown. Among 920 reproducibly identified proteins detected, 74 proteins were differentially expressed following DDX3Y siRNA treatment compared to mock treated cells. Functional grouping indicated these proteins were associated with cell cycle, cell-to-cell signaling, apoptosis and other important networks such as RNA processing and transcription regulation. Disease-based analysis confirmed DDX3Y involvement primarily in neurological and RNA metabolism disorders. Our results confirm that MSY genes are expressed in male neuronal cells, and demonstrate that Y linked DDX3 (DDX3Y) could play a multifunctional role in neural cell development in a sexually dimorphic manner.
Project description:Human embryonic stem (hES) cells have unique features: self-renewal ability and pluripotency. They can be continuously cultured in undifferentiated state and give rise to cells and tissues of all three germ layers. Thus hES cells provide a resource not only for cell replacement therapy but also for studying human developmental biology. However, much work needs to be done to understand the molecular mechanisms responsible for the maintenance of the undifferentiatiation state and the differentiation process of human embryonic stem cells. Keywords: cell type comparison design We performed gene expression profiling on 3 separate passages from 3 different hES cell lines (WA09 (H9), TE06 (I6) and BG01v), EBs samples derived from WA09 cells at 3 different time points, and 5 types of adult cells (HUVEC, HMVEC, UASMC NHA, and LFB).
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
Project description:Human embryonic stem (hES) cells have unique features: self-renewal ability and pluripotency. They can be continuously cultured in undifferentiated state and give rise to cells and tissues of all three germ layers. Thus hES cells provide a resource not only for cell replacement therapy but also for studying human developmental biology. We aimed to identify the unique signature of miRNAs in human embryonic stem cells. Keywords: cell type comparison design We performed microRNA expression profiling on 3 passages from 2 different hES cell lines (WA09 (H9) and BG01v) and 4 passages from 1 hES cell line (TE06 (I6)), EBs samples derived from WA09 cells at 4 different time points with replicates, and 5 types of adult cells (HUVEC, HMVEC, UASMC NHA, and LFB).
Project description:As part of the Dystrophia Myotonica Biomarker Discovery Initiative (DMBDI) a dataset was obtained from 35 participants, including 31 Myotonic Dystrophy type 1 (DM1) cases and four unaffected controls. All DM1 cases in this research were heterozygous for the abnormally expanded CTG repeat. The mode of the length of the DM1 CTG expansion (Modal Allele Length, MAL) was determined by small-pool PCR of blood DNA for 35/36 patients. For this work we did not attempt to measure the repeat length from muscle, due to a very high degree of repeat instability in muscle cells, and associated difficulties in its experimental measurement. One patient refused blood donation. For each of the 35 blood-donating patients mRNA expression profiling of blood was performed using Affymetrix GeneChip™ Human Exon 1.0 ST microarray. For 28 of 36 patients a successful quadriceps muscle biopsy was obtained. The muscle tissue was mRNA profiled using the same type of microarray. In total, a complete set of samples (blood and muscle) was obtained for 27 of 36 patients; samples were given a disease staging score based on muscle impairment rating. mRNA profiling was carried out by the GeneLogic service lab (on a fee-for-service basis) using standard Affymetrix hybridisation protocol.
Project description:To elucidate the Nodal transcriptional network that governs endoderm formation, we used ChIP-Seq to identify genomic targets for SMAD2/3, SMAD3, SMAD4, FOXH1 and the active and repressive chromatin marks, H3K4me3 and H3K27me3, in human embryonic stem cells (hESCs) and derived endoderm. We demonstrate that while SMAD2/3, SMAD4 and FOXH1 target binding is highly dynamic, there is an optimal signature for driving endoderm commitment. Initially, this signature is marked by both H3K4me3 and H3K27me3 as a very broad bivalent domain in hESCs. Within the first 24 hours, at a few select promoters, SMAD2/3 accumulation coincides with H3K27me3 depletion so that these loci become selectively monovalent marked only by H3K4me3. The correlation between SMAD2/3 binding, monovalent formation and transcriptional activation suggests a mechanism by which SMAD proteins coordinate with chromatin at critical promoters to drive endoderm specification. Examination of 2 different histone modifications and 4 different transcription factor associations in 2 cell types. For transcription factor analysis, three biological replicate ChIPs were pooled from each antibody, as well as input controls, for both hESCs and derived endoderm. For histone modifications, two biological replicates for H3K4me3 and three for H3K27me3 were used.
Project description:This SuperSeries is composed of the following subset Series: GSE16678: MicroRNA expression data from differentiation of human Cyt49 ESCs into definitive endoderm in feeder-free conditions GSE16681: mRNA expression data from differentiation of human ESCs into definitive endoderm, Cyt49 on matrigel GSE16687: MicroRNA expression data from differentiation of human Cyt49 ESCs into definitive endoderm on MEF feeder layers GSE16689: MicroRNA expression data from differentiation of human H9 ESCs into definitive endoderm on MEF feeder layers Refer to individual Series