Ontology highlight
ABSTRACT:
PROVIDER: PRJNA771770 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR16362700_1.fastq.gz | Fastqsanger.gz | |||
| SRR16362700_2.fastq.gz | Fastqsanger.gz | |||
| SRR16362701_1.fastq.gz | Fastqsanger.gz | |||
| SRR16362701_2.fastq.gz | Fastqsanger.gz | |||
| SRR16362696_1.fastq.gz | Fastqsanger.gz |
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Project description:Donor variation is a prominent critical issue limiting applicability of cell-based therapies. We asked whether reference protocols for pre-clinical bone marrow stromal cell (BMSC) propagation using human platelet lysate (hPL) to replace xenogeneic fetal bovine serum (FBS) might also reduce donor variability for osteo-chondral regeneration. We therefore investigated the impact of donor variability, hPL- vs FBS driven propagation and exhaustive proliferation, on BMSC epigenome, transcriptome, phenotype, coagulation risk and osteochondral regenerative function. Polychromatic flow cytometry revealed maintained canonical fibroblastic phenotype in all cells tested. We confirmed significantly declining proliferative potential in FBS-expanded BMSC after proliferative challenge. Notably, BMSC propagation under the aegis of hPL, compared to FBS, significantly increased BMSC proliferation, created significantly different gene expression trajectories and diverging surface marker signatures, already after just one culture passage. We observed limited but measurable effects on DNA methylation irrespective of culture duration. Moreover, expansion under xenogenic serum conditions resulted in significant loss of function during 3D cartilage organoid-like disk formation and significantly increased clotting risk. Superior chondrogenic function under hPL conditions was maintained over culture duration. As an additional observation, platelet blood group and isoagglutinin levels showed phenotypic changes but only minor impact on BMSC chondrogenesis and clotting behaviour.
2022-11-04 | GSE194298 | GEO
Project description:Donor variation is a prominent critical issue limiting applicability of cell-based therapies. We asked whether reference protocols for pre-clinical bone marrow stromal cell (BMSC) propagation using human platelet lysate (hPL) to replace xenogeneic fetal bovine serum (FBS) might also reduce donor variability for osteo-chondral regeneration. We therefore investigated the impact of donor variability, hPL- vs FBS driven propagation and exhaustive proliferation, on BMSC epigenome, transcriptome, phenotype, coagulation risk and osteochondral regenerative function. Polychromatic flow cytometry revealed maintained canonical fibroblastic phenotype in all cells tested. We confirmed significantly declining proliferative potential in FBS-expanded BMSC after proliferative challenge. Notably, BMSC propagation under the aegis of hPL, compared to FBS, significantly increased BMSC proliferation, created significantly different gene expression trajectories and diverging surface marker signatures, already after just one culture passage. We observed limited but measurable effects on DNA methylation irrespective of culture duration. Moreover, expansion under xenogenic serum conditions resulted in significant loss of function during 3D cartilage organoid-like disk formation and significantly increased clotting risk. Superior chondrogenic function under hPL conditions was maintained over culture duration. As an additional observation, platelet blood group and isoagglutinin levels showed phenotypic changes but only minor impact on BMSC chondrogenesis and clotting behaviour.
2022-11-04 | GSE194295 | GEO
Project description:The proliferation, migration, and osteogenic differentiation of bone mesenchymal stem cells (BMSCs) play vital roles in maintaining bone mass and metabolism. BMSC base cell therapy has become a competitive treatment option for osteoporosis.Small are critical regulators of cellular processes. Small RNA sequencing was used to detect piRNA expression during BMSC proliferation.
2024-06-01 | GSE234060 | GEO
Project description:Background: Bone marrow stromal cells (BMSCs) have classically been cultured in media supplemented with 20% fetal bovine serum (FBS). As an alternative to FBS, pooled solvent detergent apheresis platelets, HPGF-C18, was evaluated for BMSC culture. Methods: A comparison of passage 2 BMSC growth revealed that 10% HPGF-C18 produced similar cell numbers as 20% FBS. Marrow aspirates from 5 healthy subjects were cultured for 4 passages in 10% HPGF-C18 or 20% FBS and were analyzed for proliferation, colony formation efficiency (CFE), surface marker expression, suppression of mixed lymphocyte reactions (MLRs), global gene and microRNA expression analysis. BMSC supernatant cytokine and growth factor concentrations were also compared. Results: Primary cultures of marrow aspirates in 10% HPGF-C18 and 20% FBS yielded similar numbers and CFE. After 4 passages, 10% HPGF-C18 and 20% FBS yielded similar numbers of BMSCs, surface marker expression patterns and immunosuppression effects. Gene and microRNA expression analysis revealed that BMSCs cultured under the two conditions had distinct expression profiles. Gene Set Enrichment Analysis (GSEA) revealed HPGF-C18-cultured BMSCs were enriched in metabolic processing and biosynthetic pathways; cell proliferation and cell cycle pathways; and immune response pathways. FBS-cultured BMSCs were enriched in MAPK signaling, TGF-beta signaling, cell adhesion and extracellular matrix pathways. Differently expressed microRNAs were related to the osteogenesis of BMSCs. Supernatant analysis found that HPGF-C18 BMSCs displayed higher levels of PEDF and TGFB1 and lower levels of IL6, VEGF, SDF1 and PLGF. Conclusions: Traditional measures; expansion, surface marker expression and inhibition of MLRs suggest that BMSC cultured in HPGF-C18 and FBS were similar, but analysis at the molecular level revealed many differences. BMSCs cultured in HPGF-C18 should be assessed in specific functional assays that reflect application-specific potency before substituting FBS with HPGF-C18.
2018-03-21 | GSE111495 | GEO
Project description:Human bone marrow stromal cells (BMSCs) are key elements of the hematopoietic environment and they play a central role in bone and bone marrow physiology. However, how key BMSC functions are regulated is largely unknown. We analyzed the role of the immediate early response transcription factor EGR1 as key BMSC regulator and found that EGR1 was highly expressed in prospectively-isolated primary BMSCs, downregulated upon culture, and lower in non-CFU-F-containing CD45neg BM cells. Furthermore, EGR1 expression was lower in proliferative regenerating adult and fetal primary cells compared to adult steady-state BMSCs. Accordingly, EGR1 overexpression markedly decreased BMSC proliferation but considerably improved hematopoietic stroma support function as indicated by an increased production of transplantable CD34+CD90+ hematopoietic stem cells in expansion co-cultures. The improvement of BMSC stroma support function was mediated by increased expression of hematopoietic supporting genes, such as VCAM1 and CCL28. On the other hand, EGR1 knockdown increased ROS-mediated BMSC proliferation, and clearly reduced BMSC hematopoietic stroma support potential. These findings thus show that EGR1 is a key BMSC transcription factor with a dual role in regulating proliferation and hematopoietic stroma support function that is controlling a genetic program to coordinate the specific functions of BMSC in their different biological contexts.
2019-06-27 | PXD011767 | Pride
Project description:The goal of this study are to compare the transcriptome profiling (RNA-seq) of rat bone marrow mesenchymal stem cells (BMSC) transcriptome profiling (RNA-seq) under the condition of infection of different lentiviruses(shRac1 or shLuci) and/or oxygen-glucose deprivation (OGD).
2023-02-02 | GSE146261 | GEO
Project description:Human bone marrow stromal cells (BMSCs, also known as bone marrow-derived mesenchymal stem cells) are manufactured using many different methods, but little is known about the spectrum of manufacturing methods used and their effects on BMSC characteristics and function. Seven centers using, and one developing, Good Manufacturing Practices (GMP) processes were surveyed as to their production methods. Among the seven centers, all used marrow aspirates as the starting material, but no two centers used the same manufacturing methods. Two to four BMSC lots from each center were compared using global gene expression. Among the twenty-four BMSC lots from the eight centers intra-center transcriptome variability was low and similar among centers. Principal component and Unsupervised Hierarchical Clustering analysis separated all the lots from five centers into five distinct clusters. BMSCs from six of the eight centers were tested for their ability to form bone and support hematopoiesis by in vivo transplantation. Those from all six centers tested formed bone, but the quantity formed was variable highly and BMSCs from only three centers supported hematopoiesis. These results show that differences in manufacturing resulted in variable BMSC characteristics including their ability to form bone and support hematopoiesis.
2017-05-15 | GSE92640 | GEO
Project description:Bone marrow-derived mesenchymal stem cell (BMSC) is one crucial component of the multiple myeloma (MM) microenvironment and supports the malignant progression of MM. Whether BMSCs act on MM cells via exosomes has not been well characterized. Herein, we used microarrays to screen out differentially expressed miRNAs in BMSCs from patients with MM (MM-MSCs) or benign diseases (BD-MSCs).
2022-03-23 | GSE196321 | GEO
Project description:We found that mesenchymal stem cells isolated from colonic tissue express significantly increased expression of Ptgs2 or cox-2 compared to that of mesenchymal stem cells isolated from the bone marrow, one of the most common tissues from which this cell type is derived. We hypothesized that the localization of these cells in the colon exposes them to specific environmental factors that induce and/or maintain increased constitutive levels of cox-2 uniquely in this cell type. In order to identify possible candidate molecules regulating cox-2 in these cells, we decided to compare the transcriptomes of colonic mesenchymal stem cells and bone marrow-derived mesenchymal stem cells. Mesenchymal stem cells were isolated by fast plastic adherence after collagenase type I-mediated dissociation of the tissue of origin. Cells were maintained in culture in low glucose DMEM with 10mM HEPES and 10% FBS. Cells were passaged at 90-100% confluency. Approximately 4 million cells from four MSC lines, two from colon and two from bone marrow, were lysed between passages 5 and 10. Total RNA was isolated, amplified, and hybridized to Affymetrix MOE230.2 chips.
2009-12-02 | E-GEOD-19233 | biostudies-arrayexpress
Project description:Bone marrow stromal cells (BMSCs) provide hematopoietic support, immunoregulation and contain a stem cell fraction capable of skeletogenic differentiation. A heterogeneous population of primary bone marrow BMSCs were isolated from a single human donor (FH181), and a lentiviral expression system was used to overexpress human telomerase reverse transcriptase (hTERT) and generate single cell-derived immortalised BMSC lines for multi-level analysis of functional markers for BMSC subsets. All clones expressed typical BMSC cell surface antigens, however clones Y101 and Y201 displayed typical BMSC tri-lineage differentiation capacity where clones Y102 and Y202 lacked significant tri-lineage differentiation potential. High quality RNA samples were isolated from all lines, and global gene expression analysis was performed in triplicate arrays (using Agilent SurePrint G3 Human Gene Expression 8x60K v2 Microarrays) in order to identify distinguishing characteristics of these lines, compared to the parental primary BMSCs from which they were derived.
2015-07-23 | E-MTAB-3511 | biostudies-arrayexpress