Ultracentrifugation, ExoPerfectTM-MU and PEG6000 for the extraction and identification of seminal plasma exosomes: A comparative study / 医学研究生学报

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 (P0.05). 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.