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Abstract
BackgroundExosomes, internal proteins, lipids, and nucleic acids coated by phospholipid bilayer membranes, are one type of small extracellular vesicles, which can mediate cell-cell communication. In recent years, exosomes have gained considerable scientific interest due to their widely applied prospect in the diagnosis and therapeutics of human and animal diseases. In this study, we describe for the first time a feasible method designed to isolate and characterize exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells.ResultsExosomes from feline plasma, urine and adipose-derived mesenchymal stem cells were successfully isolated by differential centrifugation. Quantification and sizing of exosomes were assessed by transmission electron microscopy, flow nano analysis and western blotting. Detected particles showed the normal size (30–100 nm) and morphology described for exosomes, as well as presence of the transmembrane protein (TSG101, CD9, CD63, and CD81) known as exosomal marker.ConclusionsThe results suggest that differential centrifugation is a feasible method for isolation of exosomes from different types of feline samples. Moreover, these exosomes can be used to further diagnosis and therapeutics in veterinary pre-clinical and clinical studies.
Highlights
Exosomes, internal proteins, lipids, and nucleic acids coated by phospholipid bilayer membranes, are one type of small extracellular vesicles, which can mediate cell-cell communication
Identification of adipose-derived mesenchymal stem cells (AD-Mesenchymal stem cells (MSCs)) Differentiation of AD-MSCs After induction with adipogenic medium for 14 days, AD-MSCs gradually changed from fibroblast-like cells to flattened cells, and many different sizes lipid droplets appeared in the cytoplasm
Calcium nodules appeared on the 10th day of induced differentiation and tightly packed colonies forming nodule-like structures were observed and deposition of calcium in these cells was observed by staining with alizarin red (Fig. 2A)
Summary
Internal proteins, lipids, and nucleic acids coated by phospholipid bilayer membranes, are one type of small extracellular vesicles, which can mediate cell-cell communication. Plasmaderived exosomes (Plasma-exo), which are simple collected, have no adverse effects on health, are considered diagnostic markers for several diseases such as oncology [12], hematonosis [13], angiocardiopathy [14] or ischemic disease [15]. The study of their content (protein or nucleic acid) components is helpful for the treatment of diseases. The changes of urinary exosome-derived miRNAs and proteins can be used as
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