A study published in Cancer Cell International finds that small extracellular vesicles-educated hepatocytes in colorectal cancer may have an active role in the early stage of liver metastasis formation.
] with minor modifications. The tissue pieces were gently sliced into small fragments and incubated with collagenase D and DNase I dissolved in RPMI plain medium for 30 min at 37 °C. After incubation, the samples were passed through a 70 µm filter. The resulting filtered liquid was centrifuged at 300 × g for 10 min and 2,000 × g for 20 min and ultracentrifuged at 16,500 × g for 6 min to remove cells, tissue debris, and large EVs.
1. For sEVs isolated from conditioned culture medium and plasma samples, sEVs were prepared for electron microscopy studies using negative staining. Approximately 5 μl of the sEV suspension was deposited onto carbon-coated EM grids. After washing, the samples were fixed for 5 min in 1% glutaraldehyde and negatively stained with a 2% aqueous solution of phosphotungstic acid. The grids were viewed in a JEM 1400 Plus electron microscope operating at 80 kV equipped with a CCD camera.
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