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Single cell sequencing of human mesenchymal stem/stromal cells (MSCs) upon inflammatory licensing

ABSTRACT: We report changes of the transcriptome profile at the level of single MSCs in response to inflammatory trigger. This process is known as "MSC licensing" and represents a physiological counterregulatory mechanism. MSCs are a heterogenous population, which is highlighted by the according clustering.

ORGANISM(S): Homo sapiens

PROVIDER: GSE115149 | GEO | 2019/05/31

REPOSITORIES: GEO

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TGF-β1 licensed murine mesenchymal stromal cells show superior therapeutic efficacy in modulating corneal allograft immune rejection in vivo

Project description:We report a novel licensing strategy to improve the immunosuppressive capacity of MSCs. Licensing murine MSCs with TGF-β1 (TGF-β MSC) significantly improved their ability to modulate both the phenotype and secretome of inflammatory bone marrow-derived macrophages and significantly increased the numbers of regulatory T lymphocytes (Tregs) following co-culture assays. These TGF-β MSC-expanded Tregs also expressed significantly higher levels of PD-L1 and CD73, indicating enhanced suppressive potential. Detailed analysis of T lymphocyte co-cultures revealed modulation of secreted factors, most notably, elevated prostaglandin E2 (PGE2). Furthermore, TGF-β MSCs could significantly prolong rejection-free survival (69.2% acceptance rate compared to 21.4% for un-licensed MSC treated recipients) in a murine corneal allograft model. Mechanistic studies revealed that (i) therapeutic efficacy of TGF-β MSCs is Smad2/3-dependent; (ii) TGF-β MSC’s enhanced immunosuppressive capacity is contact-dependent and (iii) enhanced secretion of PGE2 (via prostaglandin EP4 receptor) by TGF-β MSCs is the predominant mediator of Treg expansion and T cell activation and is associated with corneal allograft survival. Collectively, we provide compelling evidence for the use of TGF-β1 licensing as an unconventional strategy for enhancing MSC immunosuppressive capacity.

2020-08-31 | GSE150008 | GEO

Single cell sequencing of human mesenchymal stem/stromal cells (MSCs) upon inflammatory licensing

Project description:Single cell sequencing of human mesenchymal stem/stromal cells (MSCs) upon inflammatory licensing

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Shear stress induces immunomodulatory signaling in bone marrow mesenchymal stromal cells (MSCs)

Project description:Bone marrow mesenchymal stromal cells (MSCs) regulate homeostasis and trafficking of cells of the blood lineage. In response to traumatic injury or infection, MSCs are believed to mobilize from the bone marrow, but it is largely unknown how egress into circulation impacts MSC function. Here we show that biomechanical forces associated with trafficking of MSCs from the bone marrow into the vasculature contribute uniquely to genetic signaling that reinforces MSC repression of immune cell activation. Laminar wall shear stress (LSS) typical of fluid frictional forces present on the lumen of arterioles stimulates increases in antioxidant and anti-inflammatory mediators, as well as an array of chemokines capable of immune cell recruitment. Importantly, LSS promotes a signaling cascade through COX2 that elevates prostaglandin E2 (PGE2) biosynthesis, permitting MSCs to suppress immune cell activation in the presence of inflammatory cues. Pharmacological inhibition of COX2 depleted PGE2 and impaired the ability of MSCs to block tumor necrosis factor-α (TNF-α) production, supporting a key role for PGE2 in the MSC immunomodulatory response to LSS. Preconditioning of MSCs by LSS ex vivo was an effective means of enhancing therapeutic efficacy in a rat model of traumatic brain injury, as evidenced by decreased numbers of apoptotic and M1-type activated microglia in the hippocampus and by retention of endogenous MSCs in the bone marrow. We conclude that biomechanical forces provide critical cues to MSCs residing at the vascular interface which influence MSC immunomodulatory and paracrine functions, thus providing unique opportunities for functional enhancement of MSCs used in therapeutic applications.

2017-05-23 | GSE82269 | GEO

Gene expression profiling of primary human bone marrow mesenchymal stromal cells (MSCs) cultured as 2D monolayers and 3D cellular aggregates (spheroids)

Project description:Human bone marrow mesenchymal stromal cells (MSCs) are conventionally cultured as adherent monolayers on tissue culture plastic. MSCs can also be cultured as 3D cell aggregates (spheroids). Optimised 3D conditions (60,000 MSCs cultured as a spheroid for 5 days) inhibited MSC proliferation and induced cell shrinkage in the absence of cell death. Primary human MSCs isolated from 2 donors were cultured under both monolayer (2D MSCs) and optimised 3D (3D MSCs) conditions. High quality RNA was isolated from all samples, and global gene expression analysis was performed in duplicate (using Agilent SurePrint G3 Human Gene Expression 8x60K v2 Microarrays) to identify gene expression changes in 3D compared to 2D MSC cultures.

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Epigenome analysis of normal and myelodysplastic sundrome (MDS) bone marrow derived mesenchymal stromal cells (MSCs)

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2019-09-06 | GSE119617 | GEO

Quantitative protein profiling of conditioned medium cultured with various mesenchymal stem/stromal cells (MSCs)

Project description:To compare the proteins secreted by human primary cultured MSCs and human iPSC-derived MSCs in the medium, each MSCs and human dermal fibroblasts were cultured in PRIME-XV MSC Expansion XSFM medium for 48 hours, and the cultured media were collected. Albumin and immunoglobulin were removed from the cultured media using High select top14 abundant protein depletion mini spin columns. The depleted media were precipitated by acetone and subjected to trypsin digestion followed by desalting. Samples were labeled with mTRAQ reagent. Tryptic peptides were fractionated through SCX-HILIC mix mode and analyzed by nLC-Orbitrap in DDA mode. MS data were searched against UniProt using Mascot.

2022-09-29 | PXD035358 | JPOST Repository

COLO 320 (Human colorectal cancer cell line) xenograft: Control (Xenograft without rat mesenchymal stem cells (MSCs)) vs Xenograft with rat MSCs

Project description:Transcriptional profiling of COLO 320 xenograft tumor cells comparing control COLO 320 xenograft without co-implanted rat MSCs with COLO 320 xenograft with co-implanted rat MSCs. The latter makes co-implanted MSCs visualization possible by using MSCs labeled by GFP under FACS and single cell microscopy. Two-condition experiment, COLO 320 xenograft without rat MSCs [COLO320 MSC(-)] vs. COLO 320 xenograft with rat MSCs [COLO320 MSC(+)]. Biological replicates: 1 control, 1 sample, paired xengraft tumor cells grown and harvested from the same mouse host. One replicate per array.

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Comparison of bovine undifferentiated MSCs, chondrogenic MSCs and chondrocytes in 3D culture

Project description:Bovine chondrocyte-seeded and mesenchymal stem cell (MSC)-seeded agarose were cultured for 28 days in chemically defined media containing 10 ng/mL TGF-beta3. Chondrogenic differentiated MSCs were compared to chondrocytes at this timepoint and to undifferentiated MSCs harvested at day 0. Donor-matched sets of chondrocytes and MSCs were used, with three donors total.

2010-10-02 | E-GEOD-18394 | biostudies-arrayexpress

Genome-wide analysis of transformed MSCs

Project description:Analysis of transformed MSCs at gene expression level. The aim in the present study was that overexpression of MYC in the MSCs alter gene expression of MSCs. Results provide important information of the response of overexpresion of MYCs, such as up- or down-regulated specific anabolic/catabolic cellular functions. Total RNA obtained from transformed MSCs or MSC culture cells at different passages.

2011-10-09 | E-GEOD-25296 | biostudies-arrayexpress

Comparative analysis of human bone marrow-derived MSCs and corneal stromal cells

Project description:Corneal stromal cells from six donors were used and expanded in standard conditions (FBS as a supplement) and xenofree conditions (HPL as a supplement). Bone marrow MSCs from one other donor were used as a comparative control. From the purified RNA, 10ng was used per sample to synthesize cDNA using the iScript Advanced cDNA synthesis kit (Bio-Rad Laboratories, Temse, Belgium) following the protocol supplied by the manufacturer. qRT-PCR assays were prepared in triplicate in 96-well PrimePCR plates for human MSCs (Bio-Rad Laboratories, Temse, Belgium), in which the primers come lyophilized per well. A total of 88 genes was targeted, which account for stemness, differentiation (osteo-, chondro-, adipo-, teno- and myogenesis), MSC-specific, and MSC-related genes. The thermal cycler used to perform the PCR was the CFX96 Real-Time System – C1000 Thermal Cycler (Bio-Rad Laboratories, Temse, Belgium). SsoAdvanced universal SYBR Green supermix was used to quantify double-stranded DNA. Dissociation of double-stranded DNA was assessed in melting curve analysis to determine the specificity. Three reference housekeeping genes were included in the assay plates (TBP, GAPDH and HPRT1) to normalize expression using the ΔΔCq method. The abundance of the transcripts in CS-MSC relative to the control BM-MSC was determined by calculating the relative normalized expression (RNE) or fold change. All analyses were performed using the Bio-Rad CFX Manager software. When gene expression exceeds a more than threefold change compared to BM-MSC it is deemed to be up- or downregulated.

2018-06-08 | GSE115453 | GEO

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