Effect of filtration on exosome extraction by ultracentrifugation

doi:10.3969/j.issn.1006-7795.2021.06.017

Abstract

Abstract: Objective To clarify the effect of filtration on the quantity and purity of exosomes extracted by ultracentrifugation, and to provide a reference for the current extraction methods of exosomes. Methods Healthy human plasma and lung cancer cell PC-9 / IR cell culture supernatants were collected and divided into unfiltration group (NC) and filtration group (GL), the filtration group was filtered with 0.22 μm filter. Exosomes were extracted by ultracentrifugation. The number and size of nanoparticles (exosomes) before and after filtration were detected by nanoparticle tracking analysis (NTA). The number and percentage of CD9⁺ exosomes (CD9⁺Exo) and CD63⁺ exosomes (CD63⁺Exo) before and after filtration were detected by Aminis imaging flow cytometry. Results The results of NTA showed that compared with the NC group, the number of exosomes decreased by 4.7 times after plasma filtration, and the number of exosomes decreased by 2.5 times after filtration of the culture supernatant. The results of flow cytometry showed that the number of CD9⁺Exo and CD63⁺Exo in plasma decreased by 5 and 2.6 times respectively after filtration, and the number of CD9⁺Exo and CD63⁺Exo in culture supernatant decreased by 3 and 2.3 times respectively after filtration. The difference was statistically significant (P<0.05). But the percentage of CD9⁺Exo and CD63⁺Exo did not significantly increase after filtered both in plasma and supernatant (P>0.05). Conclusion Filtration decrease the number of exosomes, but not improve the purity of exosomes.

Key words: exosomes, ultracentrifugation, filtration

CLC Number:

R446.11

Li Rong, Chen Jing, Li Chuanyun, Zhou Tong, Chen Huan, Chen Dexi, Qiao Yuebing, Li Weihua. Effect of filtration on exosome extraction by ultracentrifugation[J]. Journal of Capital Medical University, 2021, 42(6): 1007-1013.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2021.06.017

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2021/V42/I6/1007

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