Human islets contain four distinct subtypes of β cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE80780 | GEO | 2016/04/29
SECONDARY ACCESSION(S): PRJNA319969
REPOSITORIES: GEO
Similar Datasets
Project description:Human pancreatic islets of Langerhans contain five distinct endocrine cell types, each producing a characteristic hormone. The dysfunction or loss of the insulin-producing ? cells causes diabetes mellitus, a disease that harms millions. Until now, ? cells were generally regarded as a single, homogenous cell population. Here we identify four antigenically distinct subtypes of human ? cells, which we refer to as ?1-4, and which are distinguished by differential expression of ST8SIA1 and CD9. These subpopulations are always present in normal adult islets and have diverse gene expression profiles and distinct basal and glucose-stimulated insulin secretion. Importantly, the ? cell subtype distribution is profoundly altered in type 2 diabetes. These data suggest that this antigenically defined ? cell heterogeneity is functionally and likely medically relevant.
| S-EPMC4942571 | biostudies-literature
Project description:To characterize the molecular features of human and mouse nephron progenitors and stromal cells we performed intracellular labeling for SIX2 and/or MEIS1 antibodies, FACS, and RNA-seq. Mouse interstitial progenitors were isolated by Foxd1-Tdt transgenic line, FACS, and RNA-seq
2018-02-14 | GSE102230 | GEO
Project description:dendritic cells = improtant APC -different subtypes characterized by surface markers by FACS -combination of FACS and LC-MS powerful tool, especially for low cell numbers -direct sorting of cells into SP3-compatible lysis buffer -new protocol without washing steps after sort and before protein digestion
2024-07-25 | PXD031960 | JPOST Repository
Project description:Bacteria growing in biofilms are physiologically heterogeneous, due in part to their adaptation to local environmental conditions. Here, we characterized the local transcriptional responses of Pseudomonas aeruginosa growing in biofilms by using microarray analysis of isolated biofilm subpopulations. The results demonstrated that cells at the top of the biofilms had high mRNA abundances for genes involved in general metabolic functions, while mRNAs for these housekeeping genes were low in cells at the bottom of the biofilms. Selective GFP labeling showed that cells at the top of the biofilm were actively dividing. However, the dividing cells had high mRNAs levels for genes regulated by the hypoxia induced regulator, Anr. Slow-growing cells deep in the biofilms had little expression of Anr-regulated genes and may have experienced long-termanoxia. Transcripts for ribosomal proteins were primarily associated with the metabolically active cell fraction, while ribosomal RNAs were abundant throughout the biofilms, indicating that ribosomes are stably maintained even in slowly growing cells. Consistent with these results was the identification of mRNAs for ribosome hibernation factors (rmf and PA4463) at the bottom of the biofilms. A P. aeruginosa ∆rmf strain had increased uptake of the membrane integrity stain, propidium iodide. Using selective GFP labeling and cell sorting, we showed that the dividing cells were more susceptible to tobramycin and ciprofloxacin than the dormant subpopulation. The results demonstrate that in thick P. aeruginosa biofilms, cells are physiologically distinct spatially, with cells deep in the biofilm in a viable but antibiotic-tolerant slow-growth state.
2012-05-03 | GSE34762 | GEO
Project description:By combination of FACS sorting based on lipid sensitive Nile Red staining and small cell number proteomics the occurrence of population heterogeneity was analyzed. First population heterogeneity was identified by Nile Red staining and FACS in N. oceanica cells grown in nitrogen replete and nitrogen deplete media. For each condition two subpopulations(+NP3,+NP4,-NP3,-NP4) were detected and three replicates were sorted. Using the gathered proteome data proteins were identified and quantified with the MaxQuant software. Based on these quantification results a two sample t-test was performed to show significant regulation between +NP3 vs. +NP4 or -NP3 vs. -NP4. Goal was to understand the biological reasons for differences in lipid production between subpopulations.
2019-02-15 | PXD008721 | Pride
Project description:To characterize the progenitor-enriched CD44+NGFR+ cells present in the crypt and surface regions of the tonsil, we used an Illumina array system to analyze the transcriptome of these cells isolated from the tonsillar crypt and surface regions of three different individuals as well as the total epithelial (CD45-CD31-) populations obtained from two of these. To enrich for epithelial progeniors in human palatine tonsil tissue, hematopoietic and endothelial cells were depleted from freshly isolated cell suspensions derived from palatine tonsils using fluorescence-activated cell sorting (FACS). The resultant CD45-CD31- population was further fractionated into four subpopulations using antibodies against CD44 and NGFR. Based on the immunohistochemical phenotype and in vitro functional assays, CD44+NGFR+ subpopulations were identified as epithelial progenitor-enriched subsets. Microarray profiling was used to derive gene expression signatures of CD45-CD31- subsets (total epithelial-enriched) and CD45-CD31-CD44+NGFR+ subpopulations (progenitor-enriched).
2015-10-29 | E-GEOD-69767 | biostudies-arrayexpress
Project description:Acute kidney injury (AKI) is a major health burden in the United States. Macrophages have been shown to mediate AKI in murine models. Murine kidneys contain multiple subpopulations of kidney resident macrophages (KRM) that are transcriptionally and spatially distinct. We hypothesized the human kidney contains orthologous KRM subpopulations, both transcriptionally and spatially. We utilized kidney sections from donors diagnosed with mild to moderate AKI in order to increase the translatability of murine research to clinical settings. We used spatial transcriptomics and single cell RNA sequencing to identify five distinct human KRM subpopulations. Using the AddModuleScore function in Seurat, we were able to compare the murine KRM transcriptional profiles from pre and post-AKI to the human KRM subpopulations. As a result, we identified four orthologous to known murine KRM subpopulations. The remaining population was identified in the kidney cortex and expresses a profile similar to activate microglia, the resident macrophage population in the brain. Again, we used AddModuleScore to determine a set of marker genes that will allow for the identification of KRM subsets across species and injury.
2024-03-01 | GSE226533 | GEO
Project description:Fluorescence-activated cell sorting (FACS) is a sensitive and valuable technique to characterize cellular subpopulations and great advances have been made using this approach. Cells are often fixed with formaldehyde prior to the sorting process to preserve cell morphology and maintain the expression of surface molecules, as well as to ensure safety in the sorting of infected cells. It is widely recognized that formaldehyde fixation alters RNA and DNA structure and integrity, thus analyzing gene expression in these cells has been difficult. We therefore examined the effects of formaldehyde fixation on the stability and quantitation of nucleic acids in cell lines, primary leukocytes and also cells isolated from SIV-infected pigtailed macaques. We developed a method to extract RNA from fixed cells that yielded the same amount of RNA as our common method of RNA isolation from fresh cells. Quantitation of RNA by RT-qPCR in fixed cells was not always comparable with that in unfixed cells. In comparison, when RNA was measured by the probe-based NanoString system, there was no significant difference in RNA quantitation. In addition, we demonstrated that quantitation of proviral DNA in fixed cells by qPCR is comparable to that in unfixed cells when normalized by a single-copy cellular gene. These results provide a systematic procedure to quantitate gene expression in cells that have been fixed with formaldehyde and sorted by FACS. We compared RNA quantitation between unfixed and formaldehyde-fixed cells using the NanoString nCounter system. This analysis was performed using two cell lines, K562 and CEMx174, and human PBMCs. Three biological replicates were performed for each cell type. We also performed this comparison using human PBMCs sorted by FACS. For this analysis, we isolated PBMCs from two donors and performed two technical replicates for each donor sample.
2013-09-09 | E-GEOD-50690 | biostudies-arrayexpress
Project description:Fluorescence-activated cell sorting (FACS) is a sensitive and valuable technique to characterize cellular subpopulations and great advances have been made using this approach. Cells are often fixed with formaldehyde prior to the sorting process to preserve cell morphology and maintain the expression of surface molecules, as well as to ensure safety in the sorting of infected cells. It is widely recognized that formaldehyde fixation alters RNA and DNA structure and integrity, thus analyzing gene expression in these cells has been difficult. We therefore examined the effects of formaldehyde fixation on the stability and quantitation of nucleic acids in cell lines, primary leukocytes and also cells isolated from SIV-infected pigtailed macaques. We developed a method to extract RNA from fixed cells that yielded the same amount of RNA as our common method of RNA isolation from fresh cells. Quantitation of RNA by RT-qPCR in fixed cells was not always comparable with that in unfixed cells. In comparison, when RNA was measured by the probe-based NanoString system, there was no significant difference in RNA quantitation. In addition, we demonstrated that quantitation of proviral DNA in fixed cells by qPCR is comparable to that in unfixed cells when normalized by a single-copy cellular gene. These results provide a systematic procedure to quantitate gene expression in cells that have been fixed with formaldehyde and sorted by FACS.
2013-09-09 | GSE50690 | GEO
Project description:To characterize CD142+ ASPCs (Aregs) after exposure to an adipogenic cocktail we performed bulk RNA-seq (using BRB-seq) of total, CD142− and CD142+ mouse adipose stem and progenitor cells (ASPCs), sorted using four different anti-CD142 antibodies. ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+ CD34+ SCA1+ cells of the mouse subcutaneous stromal vascular fraction using FACS.
2022-07-04 | E-MTAB-10180 | biostudies-arrayexpress