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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

RNA-Seq of fibroblasts influenced by exosomes derived from melanoma cell line G-361

Project description:In this in-vitro study, we demonstrate the different effects of exosomes produced by melanoma cells on the functional properties of normal dermal fibroblasts and fibroblasts prepared from skin metastasis of cutaneous malignant melanoma (CMM). Both normal and cancer-associated fibroblast were prepared by a)DMEM with 10% EDS (control); b) DMEM with 10% Exosome-depleted serum (EDS) + 10 microg/ml G-361 exosomes (EXO_G361); c) DMEM with 10% EDS + 10 microg/ml exosomes from FBS (EXO_FBS). The cells were cultured for 72 hours. Transcriptome analysis was performed 72 hours after exosome application. Total RNA was isolated using RNeasy Micro Kit (Qiagen).

2022-12-31 | E-MTAB-10278 | biostudies-arrayexpress

Single-cell RNA-seq of fibroblasts influenced by exosomes derived from melanoma cell line G361

Project description:In this in-vitro study, we demonstrated the different effects of exosomes produced by G361 melanoma cells on the functional properties of normal dermal fibroblasts and fibroblasts prepared from skin metastasis of cutaneous malignant melanoma. Both normal and cancer-associated fibroblasts were cultivated in either DMEM with 10% EDS (exosome-depleted serum) (control) or DMEM with 10% EDS + 10 microg/ml G361 exosomes (EXO). The cells were harvested after 24 hours of cultivation.

2021-12-31 | E-MTAB-10290 | 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

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

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