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| Comparison of two different methods of extracting exosomes from mouse sera |
| SUN Youliang WANG Yutong▲ |
| Municipal Key Laboratory for Liver Protection and Regulation of Regeneration, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China |
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Abstract Objective To compare the effectiveness and practicability of extraction of serum exosomes by ultracentrifugation and purification kit, and to provide references for subsequent exosome studies. Methods Twelve C57BL/6 male mice aged 6-8 weeks were divided into two groups by random table method, with 6 mice in each group, that was the ultracentrifugation extraction group and the purification kit extraction group. Exosomes in mouse serum were isolated by two methods. The particle size and distribution of exosomes in the two groups were detected by Nano particle size and zeta potentiometer(Nano ZS90). The morphological structures of exosomes were extracted by transmission electron microscopy. Western blot was used to detect the expression level of exosome marker proteins. Ribonucleic acid (RNA) was isolated and identified from exosomes extracted by the two methods. Results The outside diameter of the proceeds of two groups was 30-150 nm, and morphology was lipid bilayer vesicle. Western blot results showed that the exosomes in both groups expressed the marker protein cluster of differentiation 63 and tumor susceptibility gene, and the difference was not statistically significant (P ＞ 0.05). Compared with the purification kit extraction group, the ultracentrifugation extraction group had a higher exosomes concentration, the difference was statistically significant (P < 0.05). Moreover, the RNA in the exosomes in the ultracentrifugation extraction group was mainly microRNA, while the RNA in the exosomes in the purification kit extraction group was mainly piRNA. Conclusion The ultracentrifugation and purification kit extraction method can both effectively extract exosomes, but the exosomes obtain different RNA components, which provides a more effective selection method for the study of exosomes.
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