| Objective: U251 cells are cultured in vitro.The exosomes are isolated and purified by differential centrifugation and kit respectively.To proving that the purified substances are exosomes.Comparing the advantages and disadvantages of the two methods.Methods: U251 cells were cultured in vitro.when observing the cell growth well,replaced the culture medium with medium without adding bovine serum.After one week,the supernatant of culture was collected in two groups.With differential centrifugation,kit purification exosome,respectively.Then,under the electron microscope,the morphology was observed by liquid phase loading method,and the exosome was confirmed.The morphological structure of the two groups was compared with each other,and compare the two methods.Results: U251 cells could secrete exosome and exosome vesicles were extracted under the method of differential centrifugation and kit.The exosome of differential centrifugation extraction by differential centrifuge is the same as that of exosome,and the electron microscope has a vesicle shape,which is a sphere,an ellipsoid,a low density area in the center and a deep dyed periphery.The diameter of the two groups was between 35 and 160 nm,the mean diameter of the measured centrifugal method was 99.33±4.21 nm,and the mean diameter of the kit was 98.08±4.13 nm.The random sampling results of the exosome diameter extracted by the two groups were statistically Sexual differences.Conclusion: Glioma cells can secrete exosome.Differential centrifugation,the kit can be separated and purified to the same morphosome structure exosome.The kit has the advantages of relatively simple operation,short time and less demand,but low extraction purity,high scientific research cost and difficult clinical application.Differential centrifugal method has many steps,long time-consuming,large sample demand,but easy to master technology,low cost of scientific research,high extraction purity,and further study of exosome and clinical application. |
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