Primary human bone marrow-derived mesenchymal stem cells (MSCs) treated with TNFa ligand
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Khalid Abnaof
PROVIDER: E-MTAB-5421 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Primary human bone marrow-derived mesenchymal stem cells (MSCs) were treated with recombinant human TGFb1 (10ng/ml) for different time points (1, 3, 7, 14, 24 hours)
2017-04-28 | E-MTAB-5420 | biostudies-arrayexpress
Project description:Mesenchymal stem cells are known to be recruited to the tumor and contribute to a pro-inflammatory environment. The aim of this experiment was to identify potential direct targets of hsa-miR-1246 in human bone marrow derived mesenchymal stem cells in the context of inflammation. For this purpose, miR-1246 mimic or miRIDIAN microRNA Mimic Negative Control #2 both purchased from GE Healthcare Dharmacon Inc. were transiently transfected with Lipofectamine® 2000 (Invitrogen AG) at a final concentration of 30nM into primary MSCs. Transfection time was 48h according to manufacturerâs protocol. Transfections were performed in biological triplicates each.
2016-12-30 | E-MTAB-5033 | biostudies-arrayexpress
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goal of this study is to analyse the whole transcriptome of MSCs stimulated with TGFα in order to comprehensively assess the genes regulated by EGFR signalling Methods: Libraries of cDNA were obtained from poly(A)-RNA fractions from not-stimulated or TGFα-stimulated MSCs and sequenced with a SOLiD 5500xl platform. Whole-transcriptome reads were aligned to the version 19 of the human genome (hg19) with the SOLiD LifeScope Genomic Analysis Software version 2.5 (Life Technologies) using the parameters recommended in the user’s manual. The number of observed counts (number of reads/gene) was normalized for the length of the transcript and the number of mapped reads (RPKM) (Reads Per Kilobase per Million of mapped reads). DESeq tool in R package was used to obtain a statistical evaluation of differential gene expression. Results: 1,640 genes resulted highly differentially regulated: 967 genes up-regulated with Fold Induction (FI)≥1.50, and 673 genes down-regulated with FI≤0.50. When highly regulated genes were categorized into enriched categories according to GO molecular function classification and KEGG pathways analysis, a large number of genes coding for potentially secreted proteins or for surface receptors resulted enriched following TGFα treatment. In particular, significant up-regulation of VEGFA, IL6, EREG, HB-EGF, LIF, NGF, NRG1, CCL19, CCL2, CCL25 and CXCL3 was observed. Conclusions: Taken together these findings suggest that EGFR activation in MSCs leads to a significant change in the expression of a wide array of genes coding for secreted proteins that can significantly enhance tumor progression by acting on several mechanisms within the tumor microenvironment mRNA profiles of MSCs starved overnight in serum free medium and treated for 1 hour with recombinant at a concentration of 10 ng/ml by deep sequencing, in quadruplicate, using SOLiD 5500xl.
2015-06-26 | E-GEOD-60560 | biostudies-arrayexpress
Project description:Conventional treatments for inflammatory bowel disease (IBD) have multiple potential side effects. Therefore, alternative treatments are desperately needed. In the present study, we proposed a new therapeutic tool for the treatment of IBD in murine pre-clinical models by using MSC-Exos. We demonstrated that the infused MSC-Exos are immunotolerated by the host, which is convenient for a future clinical application of MSC-Exos in IBD. To understand the molecular basis mediating the anticolitic benefit of MSC-Exos, we performed proteomic analysis using iTRAQ technology to detect the protein expression profiles in MSC-Exos and the corresponding supernatants from their parent cell MSCs. Proteomic analysis revealed that MSC-Exos were enriched in proteins involved in regulating multiple biological processes associated with the anti-colitic benefit of MSC-Exos. Particularly, metallothionein-2 in MSC-Exos was required for the suppression of inflammatory responses in macrophages. Taken together, MSC-Exos are critical regulators of inflammatory responses and may be promising candidates for IBD treatment.
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Project description:Recently, mesenchymal stem cells-derived microvesicles (MMVs) attract much attention as a strategy of cell-free treatment. In our study, we found that MMVs could improve the organ dysfunction during sepsis. To investigate the mechanism, we harvested MMVs from mesenchymal stem cells to perform proteomic analysis, and chondrocyte-derived microvesicles (CMVs) were used as a negative control, and PGC-1α overexpressed-MMVs were used as a positive control. A total of 5411 proteins were identified in this study, and differentially expressed (DE) proteins were further identified with a cut-off of absolute fold change >1.5 and a significance p-value <0.05. Compared with CMV group, 321 proteins were upregulated and 209 proteins were downregulated in MMVs. This study illustrated the differences between MMVs and CMVs, and provided a sight for investigating the protective effect of MMVs.
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Project description:Mesenchymal stromal cells were cultured in 3D PEG hydrogels for 7 days in the presence of serum-free media or conditioned media from a panel of breast cancer cells (MCF-7, MDA-MB-231, MDA-MB-231 lung-tropic, MDA-MB-231 brain-tropic, MDA-MB-231 bone-tropic). In all cases, the secretomes were collected after cancer cells were in serum-free media for 24h.
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Project description:Standardization of MSC manufacturing is urgently needed to facilitate comparison of clinical trial results. Here, we compare gene expression of MSC generated by the adaptation of a proprietary method for isolation and cultivation of a specific umbilical cord tissue-derived population of Mesenchymal Stromal Cells (MSCs) The adaptation focused on different stages of production, from cell isolation steps to cell culturing and preservation. The origin and quality of reagents were considered and steps for avoiding microbiological and endotoxin contamination of the final cell product were implemented. Cell isolation efficiency, MSC surface markers and genetic profiles originating from the use of different medium supplements were compared. The ATMP-compliant UCXM-BM-. product was also cryopreserved avoiding the use of DMSO, an added benefit in the use of these cells as an ATMP. Cells were analysed for expansion capacity and longevity. The final cell product was further characterised by flow cytometry, differentiation potential, and tested for contaminants at various passages. Gene expression profiles, genetic stability and immune properties were analysed. human umbilical cord tissue-derived MSCs (UCXM-BM-.) were isolated using clinical grade enzymes and cultivated in flask or bags using M-NM-1-MEM basal medium with 1 g/L glucose and 2 mM glutamine supplemented with either FBS,human serum or human platelet lysate.
2013-12-16 | E-GEOD-51869 | biostudies-arrayexpress
Project description:To obtain genetic evidence about the interaction between MSCs and metastatic NB cancer cells in bone marrow (BM), we isolated bone marrow-derived MSCs from children with NB and compared their global expression patterns with MSCs obtained from normal pediatric donors using the Agilent 44K microarrays. A total of eight different donors (four NB patients and four healthy donors) were analyzed and three independent microarrays hybridizations were performed for each donor.
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Project description:Chronic inflammation leading to pro-inflammatory macrophage infiltration contributes to the pathogenesis of type 2 diabetes and subsequently the development of diabetic nephropathy. Mesenchymal stem cells (MSCs) possess unique immunomodulatory and cytoprotective properties making them an ideal candidate for therapeutic intervention We used microarrays to detail changes in the gene expression profile of monocytes isolated from type 2 diabetic patients with end-stage renal disease and non-diabetic control subjects following co-culture with MSCs. Control blood samples were obtained from 4 donors from the Australian Red Cross Blood Service. Blood was also obtained from 5 diabetic patients with end-stage renal disease receiving haemodialysis at the Monash Medial Centre. CD14+ monocytes were isolated from blood using microbeads and co-cultured for 48 hours with and without human bone marrow-derived MSCs using an in vitro transwell system. An Affymetrix GeneChip Human Gene 2.0 ST array was used.
2015-11-10 | E-GEOD-65561 | biostudies-arrayexpress
Project description:MSCs comprise several percent of pluripotent-like cells named Multilineage-differentiating stress enduring (Muse) cells that express pluripotent markers at moderate levels, are collectable as cells positive for the pluripotent surface marker SSEA-3, are able to differentiate into triploblastic lineage cells, and self-renew at the single cell level (Kuroda et al, PNAS, 2010; Wakao et al, PNAS, 2011). Importantly, MSCs also comprise cells other than SSEA-3(+)-Muse cells, namely multipotent SSEA-3(-)-non-Muse MSCs that correspond to ~98% of the total MSC population. These SSEA-3(-)-non-Muse MSCs exhibit the same properties as conventional MSCs, although the non-Muse MSCs are multipotent, they are not pluripotent. In the present study, to clarify the key molecules that characterize pluripotent-like vs multipotent somatic stem cells, we separated human MSCs into SSEA-3(+)-Muse cells and SSEA-3(-)-non Muse MSCs, and analyzed both populations by scRNA-seq. SSEA-3(+) Muse cells and SSEA-3(-) non-Muse MSCs were sorted from human BM-MSCs. Sequencing libraries were prepared using Chromium single cell 3' Kit v3 (10x Genomics, Pleasanton, CA, USA) and sequenced on HiSeq2500 (Illumina, San Diego, CA, USA). Transcripts were mapped with CellRanger pipeline v3 (10x Genomics). Library construction, sequencing, and initial analysis were performed by GENEWIZ (South Plainfield, NJ, USA).
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