Comparative analysis of human bone marrow-derived MSCs and corneal stromal cells
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PROVIDER: PRJNA475096 | ENA |
REPOSITORIES: ENA
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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
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
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.
2015-09-30 | E-MTAB-3795 | biostudies-arrayexpress
Project description:Cell culture confluency affects gene expression profiles of human bone marrow-derived mesenchymal stromal cells (MSCs). High-confluency MSCs are enriched in pathways that confer them a more immunosuppressive phenotype as compared to low-confluency MSCs.
2023-08-24 | GSE231354 | GEO
Project description:The efficient clearance of dead and dying cells, also known as efferocytosis, is critical to maintain tissue homeostasis. In the bone marrow microenvironment (BMME), this role is primarily fulfilled by professional bone marrow macrophages, but recent work has shown that mesenchymal stromal cells (MSCs) act as a non-professional phagocyte within the BMME. However, little is known about the mechanism and impact of efferocytosis on MSCs in the BMME and on their function. To investigate this, we performed flow cytometric analysis of neutrophil uptake by ST2 cells, a murine bone marrow-derived stromal cell line, and in murine primary bone marrow-derived stromal cells. Transcriptional analysis showed that MSCs possess the necessary receptors and internal processing machinery to conduct efferocytosis, with Axl and Tyro3 serving as the main receptors, while MerTK was not expressed. Moreover, the expression of these receptors was modulated by efferocytic behavior, regardless of apoptotic target. MSCs derived from human bone marrow also demonstrated efferocytic behavior, showing that MSC efferocytosis is conserved. In all MSCs, efferocytosis impaired osteoblastic differentiation. Transcriptional analysis and functional assays identified downregulation in MSC mitochondrial function upon efferocytosis. Experimentally, efferocytosis induced mitochondrial fission in MSCs. Pharmacologic inhibition of mitochondrial fission in MSCs not only decreased efferocytic activity but also rescued osteoblastic differentiation, demonstrating that efferocytosis-mediated mitochondrial remodeling plays a critical role in regulating MSC differentiation. This work describes a novel function of MSCs as non-professional phagocytes within the BMME and demonstrates that efferocytosis by MSCs plays a key role in directing mitochondrial remodeling and MSC differentiation. Efferocytosis by MSCs may therefore be a novel mechanism of dysfunction and senescence. Since our data in human MSCs show that MSC efferocytosis is conserved, the consequences of MSC efferocytosis may impact the behavior of these cells in the human skeleton, including osteoporosis in aging.
2023-07-14 | GSE223279 | GEO
Project description:Limbal stromal cells were reported to resemble mesenchymal stem cells (MSCs) with multipotential differentiation cability. However, little is known about their gene expression profiles compared to MSC derived from various sources. In this study, the gene expression profile of limbal stromal cells was compared to bone marrow, adipose stromal cells and foreskin fibroblasts. In addition, we also explored the gene expression changes of ex vivo expanded limbal stromal cells when cultured in two different systems. Expanded limbal stromal cells were divided into two groups; each cultured separately on a matrigel-coated plate in DMEM/F12 medium supplemented with bFGF and LIF and the other on a normal plate in DMEM medium supplemented with 10% fetal bovine serum (FBS). Cryopreserved bone marrow mesenchymal cells, adipose stromal cells and foreskin fibroblasts were cultured-expanded until confluent. Total RNA was extracted from all the samples and subjected to microarray experiments with an Agilent platform by using Human GE 8x60k microarrays. Data analysis was carried out with GeneSpring software. A total of 871 genes were upregulated when the limbal stromal cells were cultured in the matrigel system, whereas 58 genes were consistently differentially expressed in limbal stromal cells compared to other lineages. Besides the long intergenic non-coding RNA and unknown genes, these genes represent gene ontology for cellular components, molecular function and biological process. Samples derived from the same source were closely clustered by Hierachical clustering analysis. The limbal stromal cells have a distinct molecular signature compared to MSCs from other lineages. The culture system affected the gene expression profile of limbal stromal cells tremendously. Derived limbal stromal cells were cultured using two different methods, one with matrigel and the other with FBS. Their gene expression profiles were compared. The gene expression profile of limbal stromal cells that were cultured with FBS also was compared to the gene expression profiles of bone marrow mesenchymal stem cells, adipose stromal cells and foreskin fibroblasts.
2014-07-01 | E-GEOD-38947 | biostudies-arrayexpress
Project description:Fanconi anemia (FA) is a rare genetic disorder characterized by genomic instability, developmental defects and bone marrow failure. Homeostasis of hematopoietic stem cells (HSCs) in the bone marrow partly relies on their direct or indirect interactions with the mesenchymal stem/stromal cells (MSCs). miRNAs can play a critical role during these interactions. There is no study available so far addressing the miRNA profile of bone marrow (BM-) MSCs in FA disease state. Non-coding RNA expression profiling was performed in BM-MSCs obtained from Donors (siblings of FA patients) as well as FA patients before (preBMT) and after bone marrow transplant (postBMT) using GeneChip miRNA 2.0 Array. Quality Control (QC) was performed via Normalized Unscaled Standard Errors (NUSE) and Relative Log Expression (RLE) before further analysis.
2022-09-01 | GSE183225 | GEO
Project description:To characterize and compare XF-iMSC (XenoFree-induced Mesenchymal stem/stromal cells) and various types of MSCs (Adipose-, Bone marrow-, Unbilical cord-derived), we performed a transcriptome analysis of these MSCs
2022-06-22 | GSE206172 | GEO
Project description:Fanconi anemia (FA) is a rare genetic disorder characterized by genomic instability, developmental defects and bone marrow failure. Homeostasis of hematopoietic stem cells (HSCs) in the bone marrow partly relies on their direct or indirect interactions with the mesenchymal stem/stromal cells (MSCs). miRNAs can play a critical role during these interactions. There is no study available so far addressing the miRNA profile of bone marrow (BM-) MSCs in the FA disease state. Non-coding RNA expression profiling was performed in BM-MSCs obtained from Donors, as well as FA patients before bone marrow transplantation (preBMT) using GeneChip miRNA 2.0 array. Quality Control (QC) was performed via Normalized Unscaled Standard Errors (NUSE) and Relative Log Expression (RLE) before further analysis.
2022-09-01 | GSE183226 | GEO
Project description:Limbal stromal cells were reported to resemble mesenchymal stem cells (MSCs) with multipotential differentiation cability. However, little is known about their gene expression profiles compared to MSC derived from various sources. In this study, the gene expression profile of limbal stromal cells was compared to bone marrow, adipose stromal cells and foreskin fibroblasts. In addition, we also explored the gene expression changes of ex vivo expanded limbal stromal cells when cultured in two different systems. Expanded limbal stromal cells were divided into two groups; each cultured separately on a matrigel-coated plate in DMEM/F12 medium supplemented with bFGF and LIF and the other on a normal plate in DMEM medium supplemented with 10% fetal bovine serum (FBS). Cryopreserved bone marrow mesenchymal cells, adipose stromal cells and foreskin fibroblasts were cultured-expanded until confluent. Total RNA was extracted from all the samples and subjected to microarray experiments with an Agilent platform by using Human GE 8x60k microarrays. Data analysis was carried out with GeneSpring software. A total of 871 genes were upregulated when the limbal stromal cells were cultured in the matrigel system, whereas 58 genes were consistently differentially expressed in limbal stromal cells compared to other lineages. Besides the long intergenic non-coding RNA and unknown genes, these genes represent gene ontology for cellular components, molecular function and biological process. Samples derived from the same source were closely clustered by Hierachical clustering analysis. The limbal stromal cells have a distinct molecular signature compared to MSCs from other lineages. The culture system affected the gene expression profile of limbal stromal cells tremendously.
2014-07-01 | GSE38947 | GEO