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Mesenchymal stem cell-derived exosomes stimulate cycling quiescence and early breast cancer dormancy in bone marrow

ABSTRACT: Transwell cultures were established in 6-well plates with 0.4 micron inserts. Equal amounts (6x10e4) of BCCs and MSCs were added to the outer and inner wells, respectively, in DMEM with 10% exosome-free FCS. After 24 h, the inner wells with BCCs were discarded. The MSCs were washed with PBS and then incubated with DMEM with 2% exosome-free FCS. At 48 h, exosomes were isolated from the MSC media. The exosomes from triplicate wells were added to duplicate naïve BCCs at 2.5x105/well of 6-well plates.

ORGANISM(S): Homo sapiens

PROVIDER: GSE85341 | GEO | 2016/08/08

SECONDARY ACCESSION(S): PRJNA338200

REPOSITORIES: GEO

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Mesenchymal stem cell-derived exosomes stimulate cycling quiescence and early breast cancer dormancy in bone marrow

Project description:Transwell cultures were established in 6-well plates with 0.4 micron inserts. Equal amounts (6x10e4) of BCCs and MSCs were added to the outer and inner wells, respectively, in DMEM with 10% exosome-free FCS. After 24 h, the inner wells with BCCs were discarded. The MSCs were washed with PBS and then incubated with DMEM with 2% exosome-free FCS. At 48 h, exosomes were isolated from the MSC media. The exosomes from triplicate wells were added to duplicate naÃ¯ve BCCs at 2.5x105/well of 6-well plates. qPCR miRNA expression profiling. RNA from MSCs-Exosomes were analyzed as indicated in the summary. The PCR was run on the 7300 Real-Time PCR System using the following thermal-cycling parameters: 94 0C for 15 min, 40 cycles at 940 C for 10 sec, 550 C for 30s, 700 C for 30 sec followed by a melting curve analysis.

2016-08-08 | E-GEOD-85341 | biostudies-arrayexpress

Adipose MSC suppress breast cancer progression, metastasis and inhibit EMT pathways via exosomal-miRNAs following co-culture interaction

Project description:Introduction: Mesenchymal stem cells (MSCs) are commonly known to influence the progression of cancer due to their ability to migrate to the tumor microenvironment and interact with cancer cells. Exosomes have been found secreted by most cell types and have been shown to act as cargo carrying biomolecules including microRNA (miRNAs). Thus, understanding interaction between MSCs and cancer cells via exosomal miRNAs is crucial in determining the therapeutic role of MSC in treating breast cancer cells and relapse. Methods: Exosomes were harvested from the medium of indirect co-culture of MCF7-luminal and MDA-MB-231-basal breast cancer cells (BCCs) subtypes with adipose MSCs and profiled for miRNAs using next-generation sequencing. Results: The interaction resulted in the changes of exosomal miRNAs profiles that modulate essential signalling pathways and cell cycle arrest into dormancy via inhibition of epithelial-to-mesenchymal transition (EMT). The maintenance and induction of epithelial features in MCF7 and MDA cells have led to a positive correlation with the reduction of proliferation and metastasis as well as increased drug resistance. Overall, adipose MSCs mediated delivery of consensus miRNAs particularly miR-200 family in MCF7, miR-146a in MDA and miR-941 in both BCCs subtypes via exosomes.

2020-08-18 | GSE156380 | GEO

miRNA expression profile of cells and exosomes

Project description:We hypothesized that miRNAs in the bone maroow mesenchymal stem cells (BM-MSC)-derived exosomes contributed to the phenotype change of breast cancer cells through exosome transfer. We analyzed the miRNA expression signature in BM-MSC-derived exosomes. We compared the miRNA expression levels in exosomes between BM-MSCs and adult fibroblasts (as a control). In this study, miRNA expression including in bone-marrow mesenchymal cell (BM-MSC)-derived exosomes was examined, and compared with that of exosomes derived from adult fibroblast cells or the BM-MSC cells. In addition, miRNA expression of BM-MSC exosomes was also compared with that of breast cancer cells with or without cancer stem cell marker.

2014-07-03 | E-GEOD-58027 | biostudies-arrayexpress

miRNA expression profile of cells and exosomes

2014-07-03 | GSE58027 | GEO

Comparison of the therapeutic effects between stem cells and exosomes in primary ovarian insufficiency: As promising as cells but different persistency and dosage.

Project description:Primary ovarian insufficiency (POI) refers to the loss of ovarian function under the age of 40 and results in amenorrhea and infertility. Our previous studies have shown that transplantation of mesenchymal stem cells (MSCs) and MSC-derived exosomes in chemotherapy-induced POI mouse ovaries can reverse the POI and eventually achieve pregnancy. Based on our recent studies, MSC-derived exosomes have almost equal therapeutic potentials as transplanted MSCs. However, it is still unclear whether exosomes can completely replace MSCs in POI treatment. In this study, we induced POI in C57/BL6 mice by chemotherapy (CXT) using a standard protocol. We compared the RNA expression pattern in ovarian tissue after MSC or exosome treatment. our data suggest that a minimal 10-fold increase, and ideally 100-fold increase, in the exosome concentration is required to generate more analogous results to MSC treatment.

2023-05-30 | GSE233743 | GEO

microRNA sequencing in rat cardiac fibroblasts and fibroblast-derived exosomes

Project description:We found that cardiac fibroblasts produce and secrete exosomes. miRNA profiling and TaqMan qRT-PCR experiments identified miR-21 expression to be higher in cardiac fibroblasts compared to those of miR-21*, whereas in exosomes miR-21* expression was higher compared to miR-21. The purpose of the study was to validate these findings by miRNA sequencing in cardiac fibroblasts and fibroblasts-derived exosomes. Neonatal rat cardiac fibroblasts were cultured in DMEM + 1% exosome-depleted FBS for 48h. Conditioned medium was collected and exosomes were purified by several centrifugation and filtration steps, following ultracentrifugation. Afterwards total RNA from cardiac fibroblasts and exosomes was isolated for miRNA sequencing.

2015-12-19 | E-GEOD-76175 | biostudies-arrayexpress

Modulating Mesenchymal Stromal Cell Microenvironment Alters Exosome RNA Content and Ligament Healing Capacity

Project description:Mesenchymal stromal cells (MSCs) hold great promise in regenerative medicine due to their potential to promote tissue repair and regeneration. However, applying MSC-based cell therapy in actual clinical settings remains challenging due to issues such as immunocompatibility and cell stability. Recent shifts in therapeutic strategies have highlighted MSC-derived exosomes, small vesicles carrying various bioactive molecules, as a promising cell-free therapy to promote tissue regeneration. However, it remains largely unknown regarding the ability to customize the content of MSC-derived exosomes, how alterations in the MSC microenvironment influence exosome content, and the effects of such alterations on healing efficiency and mechanical properties in tissue repair when utilized as a therapy. In this study, we used an in vitro system of human MSC-derived exosomes and an in vivo rat ligament injury model to address these questions. We found a context-dependent correlation between exosomal and parent cell RNA content. Under native conditions, the correlation was moderate but heightened with microenvironmental changes. In vivo rat ligament injury model showed that MSC-derived exosomes increased ligament max load and stiffness. We also found that changes in the MSCs' microenvironment significantly influence the mechanical properties driven by exosome treatment. Additionally, a link was identified between altered exosomal microRNA levels and expression changes in microRNA targets in ligaments. These findings elucidate the nuanced interplay between MSCs, their exosomes, and tissue regeneration.

2024-07-17 | GSE245884 | GEO

ATAC-seq on mouse embryonic stem cells for asynchronous cells, mitotic cells and purified mitotic chromosomes.

Project description:E14Tg2a mouse embryonic stem cells (ESCs) were cultured on gelatin coated plates in KO-DMEM medium supplemented with 15% FCS and 1000 U/ml leukaemia inhibitory factor. Rapidly dividing cultures of ESCs were arrested in metaphase using demecolcine (0.1 μg/ml for 6 h), achieving samples where most (85-90%) cells were in M-phase as judged by propidium iodide labelling. Condensed chromosomes were released using polyamine buffer, stained with Hoechst 33258 and Chromomycin A3 and purified by flow cytometry using a Becton Dickinson Influx equipped with specialised air-cooled lasers.

2020-06-14 | GSE147552 | GEO

Breast cancer cell line panel stimulated with estradiol

Project description:A panel of twelve breast cancer cell lines (ATCC) were exposed to 0, 1 or 10 nM 17β-estradiol (E2) for 16 hours. Cells were maintained in DMEM/F12 1:1 (DF) supplemented with 10% or 20% fetal calf serum (FCS) or 20% FCS and 0.01 mg/ml insulin. Two days before experiments, cells were seeded in 60 mm plates in DF with charcoal stripped phenol red-free serum supplemented with non-essential amino-acids and pen/strep, at a density such that 75%-90% confluence was obtained by the time of harvesting. Medium was refreshed 24 hours before exposure to 1 nM E2, 10 nM E2 or DMSO (control) for 16h. Subsequently cells were harvested and RNA was extracted using the Nucleospin RNA isolation kit (Macherey-Nagel).

2019-06-01 | GSE127760 | GEO

Inner ear exosomes and their potential use as biomarkers

Project description:Exosomes are nanovesicles involved in intercellular communications. They are released by a variety of cell types; however, their presence in the inner ear has not been described in the literature. The aims of this study were to determine if exosomes are present in the inner ear and, if present, characterize the changes in their protein content in response to ototoxic stress. In this laboratory investigation, inner ear explants of 5-day-old Wistar rats were cultured and treated with either cisplatin or gengentamicin. Exosomes were isolated using ExoQuick, serial centrifugation, and mini-column methods. Confirmation and characterization of exosomes was carried out using transmission electron microscopy (TEM), ZetaView, BCA protein analysis, and proteomics. Vesicles with a typical size distribution for exosomes were observed using TEM and ZetaView. Proteomic analysis detected typical exosome markers and markers for the organ of Corti. There was a statistically significant reduction in the exosome protein level and number of particles per cubic centimeter when the samples were exposed to ototoxic stress. Proteomic analysis also detected clear differences in protein expression when ototoxic medications were introduced. This is the first report describing exosomes derived from the inner ear. Because these exosomes varied in number and protein composition when the inner ear was exposed to ototoxic stress, the exciting possibility exists that they might be used as biomarkers to monitor inner ear function.

2018-07-04 | PXD009483 | Pride

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