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.
Project description:Exosomes are cell-derived vesicles that were found in many biological fluids such as blood, urine, and cultured medium. Exosomes are small vesicles (approximately 100nm in diameter) that contain many functional molecules like cytokines, receptors and regulating RNAs. In this study, we found that uMSC-derived exosomes accelerates wound healing and reduce myoblast formation in vivo. In vitro study showed uMSC-exosomes specific microRNAs take major roles in inhibiting myoblast differentiation of fibroblast. MicroRNA sequencing of both uMSC- and HEK293T-derived exosomes revealed significant differences between these exosomes. Further infomatic and functional analysis showed that uMSC-exosomes specific microRNA-21, -23,-145, and -125b can target different components of TGF-β signaling which attenuates the expression of α-SMA in fibroblasts, thus inhibiting myoblast differentiation
Project description:In order to establish a rat embryonic stem cell transcriptome, mRNA from rESC cell line DAc8, the first male germline competent rat ESC line to be described and the first to be used to generate a knockout rat model was characterized using RNA sequencing (RNA-seq) analysis.
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.
Project description:Fibroblasts are the powerhouses responsible for the production and assembly of extracellular matrix. Their activity needs to be tightly controlled especially within the musculoskeletal system, where changes to extracellular matrix composition affect force transmission and mechanical loading that are required for effective movement of the body. Extracellular vesicles, including exosomes, are a mode of cell-cell communication within and between tissues, which has been largely characterised in cancer. However, it is unclear what the role of healthy fibroblast-derived extracellular vesicles is during tissue homeostasis. Here, we performed proteomic analysis of exosomes derived from primary human muscle and tendon cells to identify the potential functions of healthy fibroblast-derived exosomes. Mass spectrometry-based proteomics revealed comprehensive profiles for exosomes released from healthy human fibroblasts from different tissues. We found that fibroblast-derived exosomes were more similar than exosomes from differentiating myoblasts, but there were significant differences between tendon fibroblasts and muscle fibroblasts exosomes. Exosomes from tendon fibroblasts contain higher levels of proteins that support extracellular matrix synthesis, including TGFβ1, and muscle fibroblast exosomes contain a higher abundance of MMP2. Our data demonstrates a marked heterogeneity among healthy fibroblast-derived exosomes, indicating shared tasks between exosomes of skeletal muscle myoblasts and fibroblast, whereas tendon fibroblast exosomes has a marked fibrotic potential in human tendon tissue. These findings suggest an important role for exosomes in tissue homeostasis of both tendon and skeletal muscle in humans.