Recently, Dr. YANG Huang-Tian's group from Shanghai Institute of Nutrition and Health (SINH) of the Chinese Academy of Sciences (CAS) published a detailed protocol for how to isolate and characterize the extracellular vesicles (EVs) secreted from human pluripotent stem cell-derived cardiovascular progenitor cells (hCVPC-EVs) in Extracellular Vesicles in Diagnosis and Therapy, one book of the Method in Molecular Biology series, on April 26^th, 2022.

Myocardial infarction (MI) and resulting heart failure (HF) are major causes of morbidity and mortality worldwide. Previous study by Dr. YANG’s group revealed that hCVPCs promote cardiac infarct healing when given during the early phase of MI in mouse and nonhuman primate models. It was characterized by the significant improvement in the recovery of left ventricular function and the amelioration of cardiomyocyte death as well as scar size. More recently, they found that the beneficial effects of these cells to the infarcted hearts are related to the hCVPC-EVs.

To fully achieve their values, it is essential to establish an efficient method for the isolation of hCVPC-EVs. Different isolation methods would affect the purity, activity, biological characteristics, and application efficacy of EVs.

After a comparison of various methods for EV isolation, researchers from Dr. YANG’s group developed protocols for the efficient isolation and characterization of EVs from the conditioned medium of hCVPCs , with confirmation of their therapeutic efficacy.

The approaches include collection of conditioned medium of hCVPCs followed by differential ultracentrifugation and the characterization of their properties. The isolated hCVPC-EVs display a double membrane-bound cup shape with the mode size around 110–118nm. Traditional EV marker proteins, such as Alix, TSG101, CD63, CD9, and CD81, are detected in the hCVPC-EVs.

Compared with other methods, such as the kit isolation, YANG’s group found that the protocol has the advantages of fast, simple, high EV yield, and capable of large volume of starting material. They also demonstrated that the EVs obtained via this protocol are suitable for a series of in vitro and in vivo studies, and can effectively promote cardiac repair of infarcted hearts following intramyocardial injection. Hence, this work provided a useful method for obtaining high-yield and high-quality of EVs.

Extracellular Vesicles in Diagnosis and Therapy was edited by Maurizio FEDERICO and Barbara RIDOLFI. The first part of the book is the EV Detection and Isolation; the second part is Isolation and Characterization of Tissue and Biofluid-Specific EVs; the third part is Analysis of EV Components; and the last part is EV Engineering. Overall, this book provides exhaustive pictures of current methods to detect, isolate, analyze, and modify EVs from various sources. The knowledge would be of great utility for the researchers and industry people in the EV studies and application.

Prof. YANG Huang-Tian is the corresponding author of this protocol, and Dr. WU Qiang is the first author. This work was supported by the grants from National Key R&D Program of China, the Strategic Priority Research Program of the CAS, National Natural Science Foundation of China, and Major Program of Development Fund for Shanghai Zhangjiang National Innovation Demonstration Zone.

Media Contact:
WANG Jin (Ms.)
Shanghai Institute of Nutrition and Health,
Chinese Academy of Sciences
Email: sibssc@sibs.ac.cn
Web: http://english.sinh.cas.cn/