Isolation of high-purity and high-stability exosomes from ginseng

Jinwoo Jang; Haewon Jeong; Eunjae Jang; Eungpil Kim; Youngdae Yoon; Sujeong Jang; Han-Seong Jeong; Geupil Jang

doi:10.3389/fpls.2022.1064412

Isolation of high-purity and high-stability exosomes from ginseng

Front Plant Sci. 2023 Jan 12:13:1064412. doi: 10.3389/fpls.2022.1064412. eCollection 2022.

Authors

Jinwoo Jang¹, Haewon Jeong¹, Eunjae Jang², Eungpil Kim², Youngdae Yoon³, Sujeong Jang⁴, Han-Seong Jeong⁴, Geupil Jang¹

Affiliations

¹ School of Biological Sciences and Technology, Chonnam National University, Gwangju, Republic of Korea.
² Biopharmaceutical Research Center, Jeonnam Bioindustry Foundation, Hwasun, Republic of Korea.
³ Department of Environmental Health Science, Konkuk University, Seoul, Republic of Korea.
⁴ Department of Physiology, Chonnam National University Medical School, Hwasun, Republic of Korea.

Abstract

Exosomes are nano-sized extracellular vesicles that regulate cell growth and defense by delivering bioactive cellular constituents. They are a promising material for biomedical and cosmetic utilization, especially in medicinal crops such as ginseng. One main hurdle to their usage is the need for a method to isolate stable exosomes with high purity. In this study, we first tested two methods to isolate exosomes from ginseng: ultracentrifugation, the most widely used method; and the ExoQuick system, a polymer-based exosome precipitation approach. We also designed and tested a third method in which we combined ultracentrifugation and ExoQuick methods. Size distribution analysis revealed that the exosome isolation purity by the ultracentrifugation and ExoQuick methods alone were 34.1% and 59.7%, respectively, while the combination method greatly improved exosome isolation purity (83.3%). Furthermore, we found that the combination method also increases the colloidal stability of isolated ginseng exosomes, and the increase was almost double that of the ultracentrifugation method. Lastly, we showed that the combination method can also be used to isolate high-purity and high-stability exosomes from the model plant Arabidopsis. Overall, our findings indicate that the combination method is suitable to isolate high-purity and high-stability exosomes from plants including ginseng.

Keywords: ExoQuick system; colloidal stability; exosome; ginseng; ultracentrifugation.

Grants and funding

This work was carried out with the support of the New Breeding Technologies Development Program (Project No. PJ01653503), Rural Development Administration, Republic of Korea. This work was also supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry through Agricultural Machinery/Equipment Localization Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (122022-03-1-HD020), and the National Research Foundation of Korea Grant funded by the Korean Government [NRF-2022R1A2C1003615].