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Electron Microscopy-Based Comparison and Investigation of the Morphology of Exosomes Derived from Hepatocellular Carcinoma Cells Isolated at Different Centrifugal Speeds

Published online by Cambridge University Press:  13 February 2020

Jing-Huan Deng Open the ORCID record for Jing-Huan Deng [Opens in a new window] ,
Zhong-Jie Li ,
Zi-Xuan Wang ,
Ji Feng ,
Xue-Jing Huang  and
Zhi-Ming Zeng
Jing-Huan Deng*
Affiliation:
The Guangxi Colleges and Universities Key Laboratory of Prevention and Control of Highly Prevalent Diseases, School of Public Health, Guangxi Medical University, 22 Shuangyong Road, Nanning, Guangxi Zhuang Autonomous Region530021, China
Zhong-Jie Li
Affiliation:
Department of Toxicology, School of Public Health, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region530021, China
Zi-Xuan Wang
Affiliation:
Department of Toxicology, School of Public Health, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region530021, China
Ji Feng
Affiliation:
Department of Toxicology, School of Public Health, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region530021, China
Xue-Jing Huang
Affiliation:
Department of Environmental Hygiene, School of Public Health, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region530021, China
Zhi-Ming Zeng
Affiliation:
Department of Medical Oncology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region530021, China
*
*Author for correspondence: Jing-Huan Deng, E-mail: 502696499@qq.com
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Abstract

Exosomes derived from hepatocellular carcinoma (HCC) cells are nanovesicles and are involved in the occurrence and development of HCC, they also serve as important carriers and drug targets of nanodrug delivery systems. The external shape and internal structure of exosomes are important indexes of identification, and isolated intact morphology is crucial to biological function integrity. However, given their susceptibility to various influencing factors, the external shape and internal structure of exosomes derived from HCC cells remain incompletely studied. In this study, exosomes purified from HCC cells were isolated at different centrifugation speeds and examined via multiple electron microscopy (EM) techniques. The results demonstrate that exosomes possess a nearly spherical shape and bilipid membranous vesicle with a concave cavity structure containing electron-dense and coated vesicles, suggesting the possible existence of subpopulations of exosomes with specific functions. The exosomes isolated at ultracentrifugation (UC) speed (≥110,000×g) presented irregular and diverse external morphologies, indicating the effect on the integrity of the exosomes. Transforming growth factor signaling bioactive substances (TGF-β1, S100A8, and S100A9) can be found in exosomes by performing Western blotting, showing that the internal content is associated with metastasis of HCC. These findings show that EMelectron microscopy and UC speed can affect exosome characteristics, including external shape, internal structure, and content of bioactive substances. The electron-dense and coated vesicles that had been discovered in exosomes might become new additional morphological features, which could help to improve the interpretation of experimental results and widen our understanding of exosome morphology.

Keywords

centrifugal speedelectron microscopyexosomehepatocellular carcinomamorphology
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 26 , Issue 2 , April 2020 , pp. 310 - 318
Copyright
Copyright © Microscopy Society of America 2020

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