Ultracentrifugation, ExoPerfectTM-MU and PEG6000 for the extraction and identification of seminal plasma exosomes: A comparative study
10.16571/j.cnki.1008-8199.2019.02.009
- VernacularTitle: 不同离心方法提取及鉴定精浆外泌体的效果比较
- Author:
Wen⁃bin GUO
1
;
Wan⁃song ZHANG
1
;
Cheng YANG
1
;
Jun BIAN
1
;
Jian⁃kun YANG
1
;
Cun⁃dong LIU
1
;
Tao QI
2
;
Chun⁃yan WANG
2
Author Information
1. Department of Urology,The Third Affiliated Hospital of Southerm Medical University,Guangzhou 510630, Guangdong, China
2. Department of Laboratory Medicine,Nanfang Hospital,Southerm Medical University,Guangzhou 510515, Guangdong, China
- Publication Type:Journal Article
- Keywords:
seminal plasma;
exosomes;
ultracentrifugation;
ExoPerfectTM-MU;
PEG6000
- From:
Journal of Medical Postgraduates
2019;32(2):158-162
- CountryChina
- Language:Chinese
-
Abstract:
Objective No studies have been reported on the comparison of ultracentrifugation, ExoPerfectTM-MU and PEG6000 in extracting seminal plasma exosomes. This article aimed to compare the three methods for the extraction and identification of seminal plasma exosomes. Methods Semen samples were obtained from 30 healthy donors and randomly divided into three portions, followed by extraction of exosomes from the seminal plasma by ultracentrifugation, ExoPerfectTM-MU, and 8%PEG6000, respectively. The size of the extracted exosomes was measured with the nanoparticle tracking analyzer (NTA), their morphology observed under the transmission electron microscope (TEM), and their protein biomarkers detected by Western blot. Results Significantly higher expressions of CD63 and TSG101 were found in the exosomes extracted by ultracentrifugation than in those extracted by ExoPerfectTM-MU and 8%PEG6000 (P<0.05). NTA revealed a perfect linearity of the MODE curve and insignificant difference in the diameter of the exosomes in the ultracentrifugation, ExoPerfectTM-MU and 8%PEG6000 groups (105 vs 102 vs 115 nm, P>0.05). Compared with the 8%PEG6000 group, the ultracentrifugation and ExoPerfectTM-MU groups showed significantly higher concentrations ([11.90±1.78] vs [21.20±0.98] and [19.74±1.45]×108/mL, P<0.01) and numbers of seminal plasma exosomes under TEM (4.7±1.7 vs 7.0±1.6 and 6.0±1.6, P< 0.01). Conclusion Ultracentrifugation, ExoPerfectTM-MU and 8%PEG6000 are all capable of successful extraction and identification of seminal plasma exosomes, but the former two yield more exosomes, the latter one gives a higher purity, and ExoPerfectTM-MU is simple and convenient in operation.
