Dataset Information

NanoLC-MSMS TMT comparison of native and in vitro culture-derived neutrophils

ABSTRACT: Triplicate samples of donor-matched native neutrophils (purified through fluorescence-assisted flow cytometry using CD66b as a marker) and 18-day in vitro cultured neutrophils (derived from CD34+ cells) were analysed through TMT-based quantitative proteomics to examine possible protein abundance changes underlying neutrophil cell culture.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Neutrophil, Blood

DISEASE(S): Disease Free

SUBMITTER: Pedro Moura

LAB HEAD: Ash Toye

PROVIDER: PXD052008 | Pride | 2024-06-02

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	190620_6Plex_TMT.pdResult	Other
	190620_6Plex_TMT.pdResultView	Other
	190620_6Plex_TMT.xlsx	Xlsx
	190620_6Plex_TMT_MSMSSpectrumInfo.txt	Txt
	190620_6Plex_TMT_PS.msf	Msf

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Publications

Neutrophils cultured ex vivo from CD34<sup>+</sup> stem cells are immature and genetically tractable.

Naveh Claire A CA Roberts Kiran K Zakrzewski Przemysław P Rice Christopher M CM Ponce-Garcia Fernando M FM Fleming Kathryn K Thompson Megan M Panyapiean Nawamin N Jiang Huan H Diezmann Stephanie S Moura Pedro L PL Toye Ashley M AM Amulic Borko B

Journal of translational medicine 20240531 1

<h4>Background</h4>Neutrophils are granulocytes with essential antimicrobial effector functions and short lifespans. During infection or sterile inflammation, emergency granulopoiesis leads to release of immature neutrophils from the bone marrow, serving to boost circulating neutrophil counts. Steady state and emergency granulopoiesis are incompletely understood, partly due to a lack of genetically amenable models of neutrophil development.<h4>Methods</h4>We optimised a method for ex vivo produc ...[more]

PMID: 38822352

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Project description:The molecular interactions between the maternal environment and the developing embryo that are key for early pregnancy success are known to be influenced by factors such as maternal metabolic status. Our understanding of the mechanism(s) through which these individual nutritional stressors alter endometrial function and the in utero environment for early pregnancy success is, however, limited. Here we report, for the first time, the use of an endometrium-on-a-chip microfluidics approach to produce a multi-cellular endometrium in vitro, that was exposed to glucose and insulin concentrations associated with maternal metabolic stressors. Following isolation of endometrial cells (epithelial and stromal) from the uteri of non-pregnant cows in the early-luteal phase (Day 4-7 approximately), epithelial cells were seeded in the upper chamber of the device (4-6 104 cells/mL) and stromal cells seeded in the lower chamber (1.5-2 104 cells/mL). Three different concentration of glucose (0.5, 5.0 or 50 mM) or insulin (Vehicle, 1 or 10 ng/mL) were performed in the endometrial cells at a flow rate of 1µL/min for 72 hr to mimic the rate of secretion in vivo. Quantitative differences in the transcriptomic response of the cells and the secreted proteome of in vitro-derived uterine luminal fluid (ULF) were determined by RNA-sequencing and Tandem Mass Tagging Mass Spectrometry (TMT), respectively. Changes in maternal glucose altered 21 and 191 protein-coding genes in epithelial and stromal cells, respectively (p<0.05), with a dose-dependent quantitative change in the protein secretome (1 and 23 proteins in epithelial and stromal cells, respectively). Altering insulin concentrations resulted in limited transcriptional changes including transcripts for insulin-like binding proteins that were cell specific (5, 12, and 20) but altered the quantitative secretion of 196 proteins including those involved in extracellular matrix-receptor interaction and proteoglycan signaling in cancer. Collectively, these findings highlight one potential mechanism by which changes to maternal glucose and insulin associated with metabolic stress may alter uterine function.

Dataset Information

NanoLC-MSMS TMT comparison of native and in vitro culture-derived neutrophils

Dataset's files

Publications

Neutrophils cultured ex vivo from CD34<sup>+</sup> stem cells are immature and genetically tractable.

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OmicsDI is part of the ELIXIR infrastructure

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Dataset Information

NanoLC-MSMS TMT comparison of native and in vitro culture-derived neutrophils

Dataset's files

Publications

Neutrophils cultured ex vivo from CD34&lt;sup&gt;+&lt;/sup&gt; stem cells are immature and genetically tractable.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Neutrophils cultured ex vivo from CD34<sup>+</sup> stem cells are immature and genetically tractable.