The incorporation of extracellular vesicles from MSC into HSC increase their clonogenic capacity and engraftment ability
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PROVIDER: PRJNA494576 | ENA |
REPOSITORIES: ENA
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Project description:Treatment of graft failure after allogeneic stem cell transplantation with mesenchymal stromal cells (MSC) is being evaluated clinically. MSC can exert their functions by the release of extracellular vesicles (EV). Our aim is to analyze changes induced in hematopoietic stem cells (HSC) after the incorporation of MSC-EV. MSC were isolated from healthy donors. MSC-EV were isolated by ultracentrifugation and characterized by flow cytometry (FC), Western-blot, electron microscopy and Nanosight. EV incorporation into HSC was analyzed by FC and confocal microscopy. To study HSC gene expression, RT-PCR and arrays were performed. Apoptosis and cell cycle were evaluated by FC and clonal growth by clonogenic assays in methylcellulose. The engraftment capacity of HSC was analyzed four weeks after transplantation in NOD-SCID mice. Our results showed that MSC-EV presented the characteristic morphology, size and inmunophenotype and were able to be incorporated into HSC. MSC-EV incorporation induced an a down-regulation of pro-apoptotic genes, an overexpression of genes involved in colony formation and cell proliferation together with an activation of the JAK-STAT pathway in HSC. A significant decrease in apoptosis (p=0.001), an increase of the percentage of cells in phase S and an increased CD44 expression (p=0.013) were confirmed by FC in HSC with MSC-EV. In addition, these HSC displayed a higher CFU-GM clonogenic potential (p=0.010). Finally, in the xenotransplantation model, the engraftment ability of human HSC with MSC-EV was significantly higher (p= 0.027). In summary, the incorporation of MSC-EV induces genomic and functional changes in HSC, increasing their clonogenic capacity and their engraftment ability.
2020-10-31 | GSE120803 | GEO
Project description:Engraftment and differentiation of donor hematopoietic stem cells is decisive for the clinical success of allogeneic stem cell transplantation (alloSCT) and depends on the recipient’s bone marrow (BM) niche. Presumably, a damaged niche contributes to poor graft function post-alloSCT, but the underlying mechanisms and specifically the role of BM multipotent mesenchymal stroma cells (MSC) are ill defined. Upon multivariate analysis in a cohort of 732 individuals, we observed a reduced presence of proliferation-capable MSC in BM-aspirates from patients (N=196) who had undergone alloSCT (OR 0.61, p=.028). This was confirmed by paired analysis in 30 patients showing a higher frequency of samples with a lack of MSC presence post-alloSCT compared to pre-alloSCT. This reduced MSC presence was associated with reduced survival of patients following alloSCT and specifically with impaired graft function. Post-alloSCT MSC showed diminished in vitro proliferation along with a transcriptional antiproliferative signature, upregulation of epithelial-mesenchymal transition and extracellular matrix pathways and abnormal cytokine release in coculture with hematopoietic cells. In order to avoid in-vitro-culture bias we isolated the CD146+/CD45-/HLA-DR- BM cell fraction, which comprised the entire MSC population. The post-alloSCT isolated native CD146+MSC showed a similar reduction in proliferation capacity and shared the same antiproliferative transcriptomic signature as for post-alloSCT adherence-isolated MSC. Taken together, our data show that alloSCT confers damage to the proliferative capacity of native MSC, which is associated with reduced patient survival after alloSCT and impaired engraftment of allogeneic hematopoiesis. These data represent the basis to elucidate mechanisms of BM-niche reconstitution post-alloSCT and its therapeutic manipulation.
2023-05-01 | GSE203246 | GEO
Project description:The bone marrow harbours multipotent mesenchymal stromal cells (MSCs) that nurture haematopoietic stem cells (HSCs). The extracellular matrix (ECM) is an integral part of the bone marrow, and the aim of this study was therefore to (a) examine the effect of engineered ECM substrates on MSC gene expression over time, and (b) to determine the functional ability of ECM-cultured MSCs to support HSCs. ECMs were surface immobilised using films of maleic anhydride to covalently immobilise tropocollagen or fibrillar collagen type I to the substrate. Where indicated, collagen type 1 fibrils were supplemented with heparin or hyaluronic acid. All surfaces maintained MSC viability, though cell expansion was ECM dependent. Microarray analysis indicated that both time in culture and culture substrate were important factors regulating gene expression. Surprisingly, a number of MSC genes critical for the support of HSC self-renewal were down regulated during the study period. The functional effects of the different substrates were quantitatively assessed by determining cobblestone area-forming cell frequencies and colony-forming units (CFUs). Progenitor frequencies were similar for all substrates during the first 3 weeks in culture, but thereafter the reference material, plasma-treated polystyrene (PTP), outperformed ECM-based culture substrates, as predicted from expression studies. Likewise, PTP was able to maintain CFUs for extended periods, whereas HSCs rapidly exhausted themselves when cultured on any of the ECM substrates tested. The ability to regulate the expression of stromal factors using reconstituted ECM is exciting and warrants further studies to identify the ECM components/combinations that maximise the expansion of clonogenic HSCs. Keywords: time course, human MSC, extracellular matrix
2009-08-01 | GSE12635 | GEO
Project description:Cardiac resident c-kit+ cells aroused much interest among the cardiologists at the beginning of the century. These cells are selfrenewing, clonogenic, and multipotent and possess the potential to differentiate into all cardiovascular lineages. However, one of the critical problems faced by c-kit+ cells is the poor engraftment and survival after transplantation. Therefore, researchers are focusing on developing next generation CSC products that will enhance the engraftment, survival, and regenerative capabilities of these cells. Considering this, we made a proposal that cardiac stem cells preconditioned with exosomes derived from bone marrow mesenchymal stem cells could survive and function better in a myocardial infarction model.
2015-10-01 | GSE73598 | GEO
Project description:Therapeutic use of mesangial stem cells (MSCs) for tissue repair, including heart, has great potential. MSCs from multiple sources, including those derived from human umbilical matrix, namely Wharton's jelly, may serve as a resourceful candidate. Even after more than a decade, low engraftment efficacy of MSCs, in vivo, remains a limitation which requires in-depth understanding of the mechanisms and factors involved to facilitate a better and clinically relevant outcome. We recently hypothesized and demonstrated that the sex hormone – 17-beta estradiol (E2) – facilitates protective processes within the cardiovascular system by modulating MSC function, in vitro. Here, using a proteomic approach, we investigated the angiogenic potential of MSCs in vivo and the modulatory actions of E2 on mechanisms involved in tissue repair (potentially post-ischemic injury). Employing an in vivo approach with an ovariectomized (OVX) mice model, we evaluated the effects of E2 on MSC-induced angiogenesis using Matrigel plug assay.
2022-02-17 | PXD026380 | Pride
Project description:Mesenchymal stem cells (MSC) have emerged as potent therapeutic tool for a number of pathologies, including immune ones. However, unwelcome effects of MSC on the blood coagulation were revealed in some cases, which require more in-depth analysis. In this study, we explored the trombotic properties of human MSC from umbilical cord. We revealed strong procoagulant effects of umbilical cord MSC toward human and rat whole blood and platelets-free plasma using rotational thromboelastometry and thrombodynamics tests. The similar potentiation of clotting was demonstrated for MSC-derived extracellular vesicles (EV). In order to suggest approaches to avoid unwanted effects we studied the impact of heparin supplement on MSC/EV procoagulation properties. We found that therapeutic doses of unfractionated heparin injected in the patient's blood (administered in vivo) did not abrogate the procoagulant properties of MSC. Mass-spectrometry analysis of proteins of MSC and EV involved in coagulation-associated pathways was used to evaluate mechanisms of protrombotic effects.
2019-03-26 | PXD012768 | Pride
Project description:Human mesenchymal stem cells (hMSC) play an important role in the maintenance of bone and blood. Most protocols rely on their in vitro expansion prior to clinical use. However, several groups including our own have shown that hMSC lose proliferation and differentiation ability with serial passage in culture, limiting their clinical applications. Here we show that targeting prion protein (PrP) by chemical intervention delays this process. When PrP expression was knocked down, cultures showed significant reduction in proliferation and differentiation capacity. In contrast hMSC expanded in the presence of small molecule modulator of PrP expression, 3/689, showed extended lifespan up to 10 population doublings. Upon re-plating cultures contained more than double the number of clonogenic progenitors and showed a 10 fold increase in engraftment levels in bone marrow 5 weeks post-transplant. Human MSC treated with 3/689 showed enhanced protection from DNA damage and enhanced cell cycle progression. Gene expression profiling revealed upregulation of superoxide dismutase 2 in cells treated with 3/689, dependent on PrP expression, and suggests increased scavenging of reactive oxygen species as mechanism of action. These data point to PrP as a good target to delay loss of proliferation and differentiation of hMSC with expansion in culture. Gene expression changes in hMSC expanded in the presence of 10uM small molecule modulator of prion protein 3/689 or DMSO were analysed at passage 2 and passage 8 from 3 separate donors.
2012-01-30 | E-GEOD-31205 | biostudies-arrayexpress
Project description:Expression data from hiPS lines after commitment towards mesenchymal lineage (hiPS-MSC) and expression data of mesenchymal lines (MSC), used as positive control of commitment. Following this, hiPS-MSC and MSC are seeded on scaffold to differentiate in a ligamentous (middle) and osseous (edges) part (post 21 days 3D differentiation) We used microarrays to validate efficient commitment of hiPS to generate hiPS-MSC and do whole transcriptome comparison of hiPS-MSC vs isolated MSC (positive control) and then compare the enhanced biological functions after differentiation of these cells on a scaffold
2016-12-20 | GSE58919 | GEO
Project description:Murine MSCs: MSC-exosome-hypoxia vs MSC-exosome-normoxia
| PRJNA490327 | ENA
Project description:Next Generation Sequencing of ovarian CA-MSC & MSC Transcriptomes
| PRJNA486272 | ENA