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Gene expression data of murine hematopoietic stem cells before and after in vitro culture

ABSTRACT: Gene expression data of murine hematopoietic stem cells before and after in vitro culture

PROVIDER: PRJNA681808 | ENA |

REPOSITORIES: ENA

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Gene expression data of murine hematopoietic stem cells before and after in vitro culture

Project description:Isolation of long-term reconstituting hematopoietic stem cell (HSC) has been possible by utilizing flow cytometry with a combination of multiple antibodies against cell surface markers. However those cell surface phenotypes do not represent functional HSC after in vitro culture. We compared gene expression profiling of phenotypic HSC (CD48-KSL cells) before and after in vitro culture, and discovered that cultured HSC express mast cell-related genes including Cd244. After in vitro culture, phenotypic HSC can be divided into CD244- and CD244+ subpopulations, and only CD244- cells that have low mast cell gene expression and maintain HSC-related genes sustain reconstitution potential. Induction of CD244 may partially be triggered by induction of endoplasmic reticulum (ER) stress, as chemically induced ER stress signal increased CD244+ subpopulation while ER stress suppression using a molecular chaperone, TUDCA, decreased CD244+ population, which were correlated to the output of reconstitution assay. These data suggest CD244 positivity is a potent marker to exclude non-functional HSC after in vitro culture thereby useful to elucidate mechanism of functional decline of HSC during ex vivo treatment.

2021-11-01 | GSE162400 | GEO

CD244 expression represents functional decline of murine hematopoietic stem cells after in vitro culture

Project description:CD244 expression represents functional decline of murine hematopoietic stem cells after in vitro culture

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Gene expression data of murine hematopoietic stem cells after in vitro culture and anemic stress

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Gene expression data of murine hematopoietic stem cells after in vitro culture and anemic stress

Project description:Gene expression data of murine hematopoietic stem cells after in vitro culture and anemic stress

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Molecular characterization of hematopoietic stem cells after in vitro amplification on biomimetic 3D PDMS cell culture scaffoldsMolecular characterization of hematopoietic stem cells after in vitro amplification on biomimetic 3D PDMS cell culture scaffolds

Project description:Hematopoietic stem cell transplantation (HSCT) is successfully applied since the late 1950s, however, its efficacy still needs to be improved. A promising strategy is to transplant high numbers of pluripotent hematopoietic stem cells (HSCs). Therefore, an advanced ex vivo culture system is needed that supports the proliferation and maintains the pluripotency of HSC to override possible limitations in cell numbers gained from donors. To model the natural HSC niche in vitro and thus, to amplify high numbers of undifferentiated HSCs, we used an optimized HSC cell culture medium in combination with artificial 3D bone marrow-like scaffolds made of polydimethylsiloxane (PDMS). After 14 days in vitro (DIV) cell culture, we performed transcriptome and proteome analysis of the whole cell populations. Ingenuity pathway analysis (IPA) indicated that our 3D PDMS cell culture scaffolds activated interleukin, SREBP, mTOR and FOXO signaling pathways as well as the HSC metabolism, which we confirmed by ELISA, Western blot and metabolic flux analysis. These molecular signaling pathways and HSC metabolism are well known to promote the expansion HSCs and are involved in their pluripotency maintenance. After selection and enrichment of immature CD34-positive/CD38-negative HSCs using FACS sorting, we could confirm our findings by another proteome analysis followed by IPA. Thus, we could show that our 3D bone marrow-like PDMS scaffolds activate key molecular signaling pathways to amplify the numbers of undifferentiated HSC efficiently ex vivo.

2021-11-10 | PXD025849 | Pride

Recapitulation of murine thymopoiesis in vitro from single hematopoietic stem cells

Project description:Recapitulation of murine thymopoiesis in vitro from single hematopoietic stem cells

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NanoLC-MSMS Phosphoproteomics comparison of in vitro cultured reticulocytes before and after passage through microfluidic capillaries

Project description:Triplicate samples (biological replicates) of in vitro culture-derived reticulocytes (derived from CD34+ cells) were analysed through qualitative phosphoproteomics before and after passage through a microfluidics capillary chip to investigate the mechanisms underlying shear stress recognition.

2019-09-12 | PXD013652 | Pride

in vitro murine iBC culture

Project description:The goal of this experiment is to study mouse pluripotent stem cell-derived airway basal cell (iBC) culture at a single cell resolution.

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RNA Seq of control and Cyp26b1KO HSCs after 24h of in vitro culture and treatment

Project description:The transcriptome of Ctrl and Cyp26b1KO hematopoietic stem cells (HSC) after 24h in vitro culture upon retinoid treatment was assessed by RNAseq.

2021-11-02 | E-MTAB-9752 | biostudies-arrayexpress

Proteomic profiling of FAC-sorted ex vivo and in vitro expanded Sca-1+cells derived from murine skeletal muscle

Project description:Stem cells antigen-1 (Sca-1) is an 18-kDa mouse glycosyl phosphatidylinositol-anchored cell surface protein that has widely been used for isolation of murine hematopoietic stem cells (HSCs). However, recently several studies have reported the expression of this protein in non-HSCs such as the liver, skin and muscles among others. In the current study, we used fluorescence-activated cell sorting (FACS) technique to sort Sca-1+ stem cells from mouse hindlimb muscles. The Sca-1+ cells were divided into two sets first set of Sca-1+ cells were processed for protein extraction immediately after sorting (ex vivo set) while the second set of samples, referred to as in vitro set, were expanded in cell culture after sorting and harvested after two passage and then processed for protein extraction. A high-resolution proteome map of the ex vivo and in vitro sets was generated Orbitrap Fusion Tribrid Mass Spectrometer. In total, the analysis led to the identification of 5,581 protein groups.

2021-06-21 | PXD022247 | Pride

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