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A unique population of regulatory T cells in heart potentiates cardiac protection from myocardial ischemia/reperfusion injury

ABSTRACT: Purpose: The aims of the study were to identify that transctiptome of heart Tregs was different from lymphoid organ Tregs after I/R injury. Methods: Tregs from injured hearts and paired lymphoid organs of the Foxp3-GFP transgenic mice were double-sorted by magnetic-activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) to generate RNA-sequence 3 days after myocardial ischemia/reperfusion injury (I/R injury). Results: Transcriptional profiling revealed that the transcriptome of heart Tregs has unique characteristics when compared to lymphoid Tregs.

ORGANISM(S): Mus musculus

PROVIDER: GSE155737 | GEO | 2020/11/30

REPOSITORIES: GEO

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A unique population of regulatory T cells in heart potentiates cardiac protection from myocardial infarction

Project description:Purpose: The aims of the study were to identify that transctiptome of heart Tregs was different from lymphoid organ Tregs. Methods: Tregs and conventional T cells (Tconvs) from injured hearts and paired lymphoid organs of the Foxp3-GFP transgenic mice were double-sorted by magnetic-activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) to generate RNA-sequence 7 days after myocaridal infarction (MI). Results: Transcriptional profiling revealed that the transcriptome of heart Tregs has unique characteristics when compared to lymphoid Tregs.

2020-11-30 | GSE155483 | GEO

A unique population of regulatory T cells in heart potentiates cardiac protection from myocardial infarction [scRNA-seq]

Project description:Purpose: The aims of the study were to identify that heart Tregs were clonally expanded, and single-cell TCR repertoires of heart Tregs were different from spleen Tregs. Methods: CD4+ T cells from injured hearts and paired spleens of the C57BL/6J mice were double-sorted by magnetic-activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) to generate single cell RNA-sequence and paired V(D)J sequence 7 days after myocaridal infarction (MI). Results: The characteristics of clonal restriction in heart Tregs were investigated, and there were just a little TCR-sequence sharing between spleen Tregs and heart Tregs.

2020-10-03 | GSE155619 | GEO

Purified human leukocyte subtype DNA methylation profiles

Project description:The objective of the study was to identify differentially methylated regions of DNA (DMRs) that distinguish human leukocyte subtypes, and hence serve as biomarkers for those immune cell types. This file contains Illumina Infinium HumanMethylation27 BeadChip data for human leukocyte subtypes that were purified from whole blood samples via magnetic activated cell sorting (MACS) and purity confirmed by flourescence activated cell sorting (FACS).

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Comparison of microglial proteomes of MACS and FACS-based purified CD11b+ microglia

Project description:Background Proteomic characterization of microglia has been limited by low yield and contamination by non-microglial proteins by magnetic-activated cell sorting (MACS) enrichment strategies. To determine whether a fluorescent-activated cell sorting (FACS)-based strategy overcomes these limitations, we compared microglial proteomes of MACS and FACS-based purified CD11b+ microglia in order to identify core sets of microglial proteins in adult mouse brain tissue. Results Quantitative multiplex proteomics by tandem mass tag mass spectrometry (TMT-MS) of MACS-enriched (N = 5) and FACS-purified (N = 5) adult wild-type CD11b+ microglia identified 1,791 proteins, of which 953 were differentially expressed indicating significant differences between both approaches. While the FACS-purified microglia proteome was enriched with cytosolic, endoplasmic reticulum and ribosomal proteins involved in protein metabolism and immune system functions, the MACS-enriched microglia proteome was enriched with proteins related to mitochondrial function and synaptic transmission. As compared to MACS, the FACS microglial proteome showed strong enrichment for canonical microglial proteins while neuron, astrocyte, and oligodendrocyte proteins were decreased. Interestingly, we observed enrichment of endothelial specific proteins in the FACS microglia proteome. By comparing FACS-purified microglia proteomes with transcriptomes, we observed highly concordant as well as highly discordant proteins that were abundant at the protein level but low at the transcript level. Conclusions We demonstrate that TMT-MS proteomics of FACS-purified adult microglia is superior to column-based enrichment approaches, resulting in purer and highly-enriched microglial proteomes. We also define core sets of highly-abundant adult microglial proteins including Moesin (Msn) that can guide future studies.

2020-05-11 | PXD015652 | Pride

LC-MS/MS analysis of isolated murine brain endothelial cells (EC) and full brain tissue (FT) preparations from 3-months-old mice followed by label-free quantification (LFQ)

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Project description:Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption.

2022-11-08 | PXD038027 | Pride

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Project description:Human dermal microvascular endothelial cells (HDMECs) were isolated via the magnetic-activated cell sorting (MACS) method from skin biopsies of patients with DFUs and healthy controls. We then performed gene expression profiling analysis using data obtained from RNA-seq of these cells.

2023-09-30 | GSE219078 | GEO

Intralipid protects the heart in late pregnancy against ischemia/reperfusion injury

2016-02-09 | GSE77698 | GEO

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