Analysis of globle gene expression changes observed in fibroblasts isolated via explant outgrowth and 45-minute differential binding
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PROVIDER: PRJNA558962 | ENA |
REPOSITORIES: ENA
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Project description:Fibroblasts are a ubiquitous cell type that are found in many tissue types and are associated with a variety of diseases including most fibrotic disorders. We isolated fibroblast cells using explant outgrowth methodologies and compare the genomic signature of these cells to cells isolated using 45-minute differential binding.
2019-08-07 | GSE135456 | GEO
Project description:Streptococcus sp. 45 strain:45 Genome sequencing
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Project description:In MINUTE-ChIP, native chromatin is fragmented using Micrococcal nuclease, and subsequently blunted and ligated to double-stranded DNA adaptors that include a T7 RNA polymerase promoter and a sample barcode sequence. Finally, samples are combined and subsequent ChIP reactions are performed with the pooled samples. ChIP material is prepared into an Illumina-compatible library using linear amplification by virtue of T7 RNA polymerase, reverse transcription and a low-cycle library PCR amplification. Native MINUTE-ChIP is based on Mint-ChIP, developed by the Bernstein lab (van Galen et al., 2016; PMID: 26687680). We have introduce unique molecule (UMI) counting and paired-end mapping of the chromatin fragments to this method, which we then termed MINUTE-ChIP for multiplexed indexed unique molecule T7 amplification end-to-end sequencing. Here, we generate a standard curve for H3K27me3 and demonstrate that MINUTE-ChIP has a large linear dynamic range, thus MINUTE-ChIP quantitation is proportional to real quantities.
2019-06-21 | GSE133056 | GEO
Project description:Pericytes are critical cellular components of the microvasculature that play a major role in vascular development and pathologies, yet their study has been hindered by lack of a standardized method for their isolation and growth. Here we report a method for culturing human pericytes from a readily available tissue source, placenta, and provide a thorough characterization of resultant cell populations.We developed an optimized protocol for obtaining pericytes by outgrowth from microvessel fragments recovered after enzymatic digestion of human placental tissue. We characterized outgrowth populations by immunostaining, by gene expression analysis, and by functional evaluation of cells implanted in vivo.Our approach yields human pericytes that may be serially expanded in culture and that uniformly express the cellular markers NG2, CD90, CD146, alpha-SMA, and PDGFR-beta, but lack markers of smooth muscle cells, endothelial cells, and leukocytes. When co-implanted with human endothelial cells into C.B-17 SCID/bg mice, human pericytes invest and stabilize developing human endothelial cell-lined microvessels.We conclude that our method for culturing pericytes from human placenta results in the expansion of functional pericytes that may be used to study a variety of questions related to vascular biology.
| S-EPMC2928225 | biostudies-literature
Project description:In plant axillary bud dormancy and outgrowth are regulated by phytohormones, but it is still unknown about its molecular mechanism. We reveal that Arabidopsis axillary buds located at axil of rosette leaves show dormancy and that this is broken by the decapitation of main stem, resulting in the bud outgrowth. To investigate about the molecular mechanisms of dormancy and outgrowth, we carried out gene expression analysis during axillary shoot outgrowth in Arabidopsis wild type Columbia accession. Since axillary buds did not initiate outgrowth (dormancy) at 5 day after bolting of main stem, we used 5-day bolted plants as a control (before decapitation). Then, main stems were decapitated, and axillary shoots were collected at 24 hours after decapitation (named as growing shoot). Total RNA was prepared from either control or growing shoots and used for the microarray analysis. We carried out duplicate microarray analysis using independent plant materials.Ref):Tatematsu et al., Plant Physiol. 138: 757-766 (2005). Keywords: Expression profilling by array 4 samples were used in this experiment
2009-11-17 | E-GEOD-18987 | biostudies-arrayexpress
Project description:Transcriptome and chromatin changes during CRC organoid outgrowth as proxy for metastatic outgrowth
| PRJNA853548 | ENA
Project description:Purpose: The goals of this study is to analyze the transcriptome change during retinal explant culture treated with pharmacological chaperone of rhodopsin (YC-001). Methods: The WT and RhoP23H/+ mouse eyes were enucleated at post natal day 15 and retinal explant were cultured for 24 hours followed by the 24 hours treatment with pharmacological chaperone of rhodopsin (YC-001) and dmso vehicle control. Another group of P23H/+ retinal explant was treated further till 9 days. We compare the transcript of RhoP23H/+ dmso with WT dmso at 24 hour treatment (1 days in vitro (1DIV)). RhoP23H/+ YC-001 were also compared with RhoP23H/+ dmso treated retinal explant at 1DIV and 9DIV. Results: A total of 338 differentially expressed genes (DEGs) were identified comparing the RhoP23H/+ vs. WT retinal explants with DMSO and 56 DEGs were identified comparing RhoP23H/+ YC-001 treated vs. DMSO treated retinal explant at 1 DIV. Further we observed an increased number of DEGs (8,597) comparing RhoP23H/+ vs dmso vehicle control at 9DIV. DEGs were identified using stringency with >1.5-fold change and P-value<0.05. Interestingly, we found 40 common genes when comparing RhoP23H/+ YC-001 treated vs. DMSO at 1DIV and 9DIV. Conclusions: This study used RNA-seq technology which represents the transcriptome of WT and RhoP23H/+ mouse retinal explant with and without pharmacological chaperone of rhodopsin (YC-001). Our NGS results show that YC-001 affect many biological pathways including visual transduction, protein homeostasis pathways and decreases the expression of immune response activation genes, to rescue the rhodospin homeostasis and retinal morphology in the retinal explant of P23H Rho mouse model of retinitis pigmentosa (RP).
2021-07-10 | GSE179754 | GEO
Project description:We have undertaken a genome-wide study of transcriptional activity in embryonic superior cervical ganglia (SCG) and dorsal root ganglia (DRG) during a time course of neurite outgrowth in vitro. Gene expression observed in these models likely includes both developmental gene expression patterns and regenerative responses to axotomy, which occurs as the result of tissue dissection. Comparison across both models revealed many genes with similar gene expression patterns during neurite outgrowth. These patterns were minimally affected by exposure to the potent inhibitory cue Semaphorin3A, indicating that this extrinsic cue does not exert major effects at the level of nuclear transcription. We also compared our data to several published studies of DRG and SCG gene expression in animal models of regeneration, and found the expression of a large number of genes in common between neurite outgrowth in vitro and regeneration in vivo. Experiment Overall Design: We wished to determine the transcriptional profiles of neurons undergoing neurite outgrowth in vitro. We were particularly interested in finding genes whose expression is generally associated with the process of neurite outgrowth, rather than with cell type-specific effects. Thus, in order to avoid focusing on transcripts unique to one tissue type versus another, we used a comparative strategy to look for effects that were common to two tissue types and therefore more likely to be involved in the general process of neurite outgrowth. While these explants contain multiple cell types, we felt this was preferable to the more disruptive conditions required to dissociate neurons or obtain a pure neuron population. To this end, we monitored gene expression in cultured explants from SCG and DRG using DNA microarrays. We initiated our studies by culturing embryonic day 13 (E13) mouse SCG in vitro and harvesting tissue for RNA isolation at time points from 2 to 65 hours. Time points were selected to detect both fast, short-term responses (2, 5 and 12 hours), as well as sustained, long-term changes (24, 40, and 65 hours). Samples were hybridized to Affymetrix MG-U74v2 A and B microarrays, with RNA from acutely dissected explants serving as a baseline reference. We followed these experiments with a parallel analysis of a more heterogeneous tissue type, the DRG, which is more frequently used than SCG for in vivo studies of neurite regeneration. Cervical and upper thoracic DRG from E12 embryos were cultured with NGF (the same trophic support as in SCG cultures), harvested at time points from 2 to 40 hours, and hybridized to Affymetrix MOE 430A microarrays.
2008-06-16 | E-GEOD-9738 | biostudies-arrayexpress
Project description:In plant axillary bud dormancy and outgrowth are regulated by phytohoromones, but it is still unknown about its molecular mechanism. We reveal that Arabidopsis axillary buds located at axil of rosette leaves show dormancy and that this is broken by the decapitation of main stem, resulting in the bud outgrowth. To investigate about the molecular mechanisms of dormancy and outgrowth, we carried out gene expression analysis during axillary shoot outgrowth in Arabidopsis wild type Columbia accession. Since axillary buds did not initiate outgrowth (dormancy) at 5 day after bolting of main stem, we used 5-day bolted plants as a control (before decapitation). Then, main stems were decapitated, and axillary shoots were collected at 24 hours after decapitation (named as growing shoot). Total RNA was prepared from either control or growing shoots and used for the microarray analysis. We carried out duplicate microarray analysis using independent plant materials.Ref):Tatematsu et al., Plant Physiol. 138: 757-766 (2005). Keywords: Expression profilling by array
2009-11-18 | GSE18987 | GEO
Project description:Transcriptome and chromatin changes during CRC organoid outgrowth as proxy for metastatic outgrowth [bulkRNAseq_AKPS]
| PRJNA853559 | ENA