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从人参中分离高纯度和高稳定性的外泌体。

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

作者信息

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

机构信息

School of Biological Sciences and Technology, Chonnam National University, Gwangju, Republic of Korea.

Biopharmaceutical Research Center, Jeonnam Bioindustry Foundation, Hwasun, Republic of Korea.

出版信息

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

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 . Overall, our findings indicate that the combination method is suitable to isolate high-purity and high-stability exosomes from plants including ginseng.

摘要

外泌体是纳米级的细胞外囊泡,通过传递生物活性细胞成分来调节细胞生长和防御。它们是生物医学和化妆品应用中一种很有前景的材料,尤其是在人参等药用作物中。其应用的一个主要障碍是需要一种方法来分离出高纯度的稳定外泌体。在本研究中,我们首先测试了两种从人参中分离外泌体的方法:超速离心法,这是使用最广泛的方法;以及ExoQuick系统,一种基于聚合物的外泌体沉淀方法。我们还设计并测试了第三种方法,即将超速离心法和ExoQuick方法结合起来。尺寸分布分析表明,单独使用超速离心法和ExoQuick方法分离外泌体的纯度分别为34.1%和59.7%,而结合方法大大提高了外泌体分离纯度(83.3%)。此外,我们发现结合方法还提高了分离出的人参外泌体的胶体稳定性,且提高幅度几乎是超速离心法的两倍。最后,我们表明结合方法也可用于从模式植物中分离高纯度和高稳定性的外泌体。总体而言,我们的研究结果表明,结合方法适用于从包括人参在内的植物中分离高纯度和高稳定性的外泌体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0c86/9878552/bd9c80c964c7/fpls-13-1064412-g001.jpg

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