Adult mucous neck cells from corpus gastric epithelium
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE42946 | GEO | 2013/05/31
SECONDARY ACCESSION(S): PRJNA183932
REPOSITORIES: GEO
Similar Datasets
Project description:In this experiment, mucous neck cells from the gastric epithelium of normal, adult C57/B6 mice were laser-capture microdissected to determine gene expression in neck cells relative to pit cells, parietal cells, and zymogenic cells, whose expression profiles were previously deposited in GEO. In this screening experiment, only a single replicate Mouse Genome 430 2.0 GeneChip was generated from laser-capture microdissected cells in the neck region of the gastric unit (identified by positive staining with AlexaFluor-488-GS-II lectin positivity and/or position in the middle of the gastric unit with low labeling with E-cadherin and seconndary Alexafluor-488). RNA was purified by PicoPure kit, RNA integrity verified on Agilent Bioanalyzer, and total RNA pooled from multiple dissections and multiple mice to make ~100 ng RNA, which was then amplirifed, labeled, and fragmented (by Arctureus RiboAmp HS kit followed by the RNA Amplification and Labeling Kit from Enzo Life Sciences). Biotinylated cRNA probes were hybridized to the GeneChips. Given the inevitable contamination of the RNA with RNA from parietal cells, this GeneChip is used only in comparisons with laser-captured parietal cells described in GSE5018 as a baseline reference.
2013-05-31 | E-GEOD-42946 | biostudies-arrayexpress
Project description:Continuous regeneration of digestive enzyme (zymogen) secreting chief cells is a normal aspect of stomach function that is disrupted in pre-cancerous lesions. Regulation of zymogenic cell (ZC) differentiation is poorly understood. Here we profile Parietal, Pit, and Zymogenic cells for comparison and study. Keywords: Bhlhb8, Mist1, mucous neck cell, laser-capture microdissection, zymogenic, gastric cell differentiation
2006-10-20 | GSE5018 | GEO
Project description:MIST1 is a bHLH transcription factor that is necessary for the maturation of gastric zymogenic cells as they differentiate from their precursor mucous neck cells. In this experiment, mucous neck cells and zymogenic cells of normal, adult C57BL/6 and MIST1 knockout mice were laser-capture microdissected in order to determine MIST1-dependent, zymogenic cell specific gene expression.
2013-01-12 | GSE43441 | GEO
Project description:MIST1 is a bHLH transcription factor that is necessary for the maturation of gastric zymogenic cells as they differentiate from their precursor mucous neck cells. In this experiment, mucous neck cells and zymogenic cells of normal, adult C57BL/6 and MIST1 knockout mice were laser-capture microdissected in order to determine MIST1-dependent, zymogenic cell specific gene expression. Stomachs were excised immediately following sacrifice, quickly flushed with room-temperature PBS, inflated by duodenal injection of OCT compound, frozen in Cytocool II, and cut into serial 7-M-NM-<m-thick cryosections, which were mounted on Superfrost slides, fixed in 70% EtOH, rehydrated in nuclease-free water, and then incubated in Alexa Fluor 488-conjugated Griffonia simplicifolia GS-II (diluted 1:500 in nuclease-free water) for 15 min. Sections were washed in nuclease-free water and dehydrated in graded ethanol followed by xylene. ZCs were identified as corpus cells that were basal to GS-II labeling and which did not show the dark silhouettes and characteristic shape of parietal cells following xylene dehydration. Four wild-type mice and 5 Mist1M-bM-^HM-^R/M-bM-^HM-^R mice were used for dissection (PixCell II LCM apparatus [7.5-M-NM-<m spot diameter] and CapSure HS LCM caps) to generate two caps per mouse. RNA was purified by PicoPure kit, and RNA integrity was confirmed by an Agilent 2100 Bioanalyzer. All RNA from each cap was treated with DNase I and then reverse transcribed using the SuperScript III (Invitrogen) standard protocol (most cDNA syntheses started with 10 ng of total RNA). Biotinylated cRNA probes were hybridized to the GeneChips. Gene chip arrays used in these experiments were Affymetrix Mouse Gene 1.0ST arrays. Chip quality control and GeneChip to GeneChip comparisons to generate expression profiles were performed using dChip.
2013-01-12 | E-GEOD-43441 | biostudies-arrayexpress
Project description:Continuous regeneration of digestive enzyme (zymogen) secreting chief cells is a normal aspect of stomach function that is disrupted in pre-cancerous lesions. Regulation of zymogenic cell (ZC) differentiation is poorly understood. Here we profile Parietal, Pit, and Zymogenic cells for comparison and study. Experiment Overall Design: Samples were obtained through Laser-capture microdisection of gastric epithelium. The three samples come from enrichments for Zymogenic Cells (ZC), Parietal Cells (PC), and Mucus Pit Cells (Pit). Experiment Overall Design: Individual parietal cells (visualized by DBA-positivity and autofluorescence) from well-oriented gastric units were dissected (PixCell II LCM apparatus, 7.5-µm spot diameter; CapSure HS LCM caps, Arcturus, Mountain View, CA) one-at-a-time from the pit zone and collected for GeneChips. Pit cells (E-cadherin-positive, DBA-negative) were then collected from the same gastric units. Only the pit cells in a 2-3-cell-thick region at the apex of the gastric unit â?? but not yet upon the gastric surface â?? were taken. ZCs (E-cadherin-positive, GSII/DBA-negative cells in the base zones) were collected from different slides in corpus gastric units after potentially contaminating basal parietal cells had first been dissected and discarded. 3000-5000 individual cells from each cell lineage were isolated from 4-5 individual mice, and RNA was purified by PicoPure kit (Arcturus). RNA integrity was verified, and RNAs for each lineage were pooled from multiple mice and multiple dissections to make 20 ng total RNA, which was then amplified, labeled, and fragmented (by the Arcturus RiboAmp HS kit followed by the RNA Amplification and Labeling Kit from Enzo Life Sciences). The resulting biotinylated cRNA probes were hybridized to Affymetrix (Santa Clara, CA) GeneChip® Mouse Genome 430 2.0 microarrays.
2008-06-13 | E-GEOD-5018 | biostudies-arrayexpress
Project description:A dozen of cells expressing various marker genes was defined as adult gastric epithelial stem cells. Do these cells represent different cell types or they are just the same cell with cycling expression of different marker genes? The origin of the adult stem cells could answer this question. Yet, the identity of the embryonic gastric progenitors was not known. We performed single cell RNA-sequencing to characterize cellular composition of the embryonic stomach at the time of initiation of cytodifferentiation. Our analysis of the scRNA-seq data led to identification of two different embryonic gastric epithelial progenitors. Further, we have defined signals that control their maintenance and differentiation along four main lineages, such as mucous pit, zymogenic, parietal and endocrine cells.
2020-09-04 | GSE137908 | GEO
Project description:A profile of murine gastric epithelial cells: Parietal, Zymogenic, Pit
| PRJNA95805 | ENA
Project description:Mice were infected with Helicobacter pylori strain SS1 for 2, 7, 14 or 28 days or left uninfected. They were sacrificed at the appropriate times, the stomachs were removed and one half was plated for colony counts. The other half was embedded in OCT and cryosections were produced for 6-7 animals per timepoint. The sections were stained using the Histogene dye and roughly 500-1000 cells per gastric epithelial cell type were harvested by laser microdissection. RNA was prepared from these samples, subjected to two rounds of amplification, labelled and hybridized to 40.000 element murine cDNA arrays. The array names are composed as follows: "LCM" stands for laser capture microdissection; "mock" or "SS" stands for mock-infected or SS1-infected, respectively; "2,7,14 and 28" refers to the length of infection; "1-4" corresponds to the numbers given to every animal.The cellular origin of the RNA is represented by "mucus producing" or pit cell, "parietal" or acid-producing cell and "chief" or zymogenic cell. Between 5 and 7 samples were harvested per timepoint and cell type. Where the first labelling reaction and hybridization did not produce sufficiently good data (less than half of all spots made a regression correlation cutoff of >0.6), both were repeated (indicated by the addition of "relabeled" to the array name). The differential regulation of genes can be analyzed in both a time and cell type specific manner using publicly available software. Cell Type: "mucus producing" or pit cell (pit), acid-producing cell (parietal) and zymogenic cell (chief) time_series_design
2012-05-11 | E-GEOD-37938 | biostudies-arrayexpress
Project description:Mice were infected with Helicobacter pylori strain SS1 for 2, 7, 14 or 28 days or left uninfected. They were sacrificed at the appropriate times, the stomachs were removed and one half was plated for colony counts. The other half was embedded in OCT and cryosections were produced for 6-7 animals per timepoint. The sections were stained using the Histogene dye and roughly 500-1000 cells per gastric epithelial cell type were harvested by laser microdissection. RNA was prepared from these samples, subjected to two rounds of amplification, labelled and hybridized to 40.000 element murine cDNA arrays. The array names are composed as follows: "LCM" stands for laser capture microdissection; "mock" or "SS" stands for mock-infected or SS1-infected, respectively; "2,7,14 and 28" refers to the length of infection; "1-4" corresponds to the numbers given to every animal.The cellular origin of the RNA is represented by "mucus producing" or pit cell, "parietal" or acid-producing cell and "chief" or zymogenic cell. Between 5 and 7 samples were harvested per timepoint and cell type. Where the first labelling reaction and hybridization did not produce sufficiently good data (less than half of all spots made a regression correlation cutoff of >0.6), both were repeated (indicated by the addition of "relabeled" to the array name). The differential regulation of genes can be analyzed in both a time and cell type specific manner using publicly available software. Cell Type: "mucus producing" or pit cell (pit), acid-producing cell (parietal) and zymogenic cell (chief)
2012-05-12 | GSE37938 | GEO
Project description:Gastric ulcer, which affect many of patients and is deeply related with gastric cancer, is caused by chronic gastric acid stimulation. Stomach fundus, the main body of stomach, is a major source of gastric acid and peptidase for food digestion. Recapturing the main body of stomach requires mainly 3 functionally differentiated cells; parietal (oxyntic) cells, chief (zymogenic) cells, and surface mucous foveolar (pit) cells. We have previously shown the induction of stomach tissue with functional secreting activities by directed differentiation of mouse embryonic stem cells (ES cells) to stomach primordium with both gut epithelium and splanchnic mesoderm. However, generating human stomach with fundus and such functions has been elucidated and a long-desired goal. Here, we describe the method for establishing human embryonic stem cell-derived stomach organoids with fundus gland structure. Along with mouse stomach development and de novo stomach generation from mouse ES cells in vitro, we observed gut-like structure formation from human embryonic stem cells by induction of both endoderm and mesoderm. These human embryonic gut could differentiate into stomach primordium by growth factor stimulation as well as stomach development, and form stomach tissue in three-dimensional organoid culture. Furthermore, these stomach organoids contain fundus-like gland with parietal cells and chief cells, some of secreting activities, and is transcriptionally close to human stomach. Human functional stomach derived from embryonic stem cells represent powerful tools for analying human stomach development, and gastric ulcer related disease including gastric tumorgenesis.
2017-08-31 | GSE79673 | GEO