Agent for modulating the dissemination of cancer cells, product comprising said agent, and use of said agent in the treatment and/or prevention of cancer
A medicament and application technology, applied in the field of medicaments regulating the spread of cancer cells, especially metastatic cancer cells, can solve problems such as low efficiency and lethality
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[0116] Materials and Methods
[0117] Purification and Characterization of Exosomes
[0118] Exosomes were isolated and purified by ultracentrifugation from the ascites of patients diagnosed with ovarian cancer at the Campustral University Hospital in Santiago. Briefly, 36 ml of ascitic fluid was centrifuged sequentially at 300 g for 10 min, 2,000 g for 20 min, and 10,000 g for 30 min in order to collect cell pellets and debris. The supernatant was filtered through a syringe filter with a pore size of 22 μm, and then centrifuged at 100,000 g for 90 min (SW80 rotor; Beckman Instruments). The collected cell mass was resuspended in 100 μL of phosphate buffered saline, aliquoted and stored at -20°C. Exosomes were quantified by solubilizing exosomal proteins and using nanodroplet and Bradford methods. The size of exosomes was analyzed by 90° angle detection and laser Doppler anemometry and anemometry (Zetasizer 3000HS Malvern Instruments), respectively, at 25°C. Exosome morphol...
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