Comparison of two methods for extracting exosomes from the nucleus accumbens in mice

Abstract

Background: Exosomes bind to and are endocytosed by neurons of various brain regions. Methods for isolating and extracting exosomes from specific brain samples are critical. At present, the most important extractive methods for exosomes are Ultracentrifugation and exosome isolation kit extraction. Both of these extraction methods have applications in neuroscience. We compare these methods to reveal the differences.

Methods: We sectioned the nucleus accumbens of mice, and isolated exosomes. A culture medium containing exosomes was extracted using ultracentrifugation (UC) and a total exosome isolation kit (TEI). The exosomes were examined using transmission electron microscopy (TEM), measurement regarding the diameter of the exosomes was done, and the thermal allodynia and western blotting analysis were also conducted, respectively.

Results: Transmission electron microscopy observations showed that the ultracentrifugation samples had higher purity and fewer impurities than the kit samples. The results from the two methods were then compared with a number ratio regarding the percentage was not statistically significant. Marker protein tests showed that proteins were expressed under both methods. The thermal allodynia testing observed that the two extraction methods did not affect pain behavior regarding the detection. After the kit extraction method, there were substantial white subjects suspended by PBS.

Conclusion: Our study compared the different protocols regarding exosome extraction from the nucleus accumbens and compared the quality of two principal methods for exosome extraction from a culture medium containing exosomes. It was found that the extraction quality of exosomes by ultracentrifugation was better, but the technical difficulty was greater.

Keywords: CCI; Exosomes; Kit method; Nucleus accumbens; Pain; Ultracentrifugation.