@inbook{1f0804c2fc314f49a673b4f23029e073,
title = "Comprehensive Method for Exosome Isolation and Proteome Analysis for Detection of CCN Factors in/on Exosomes",
abstract = "Cellular Communication Network (CCN) proteins are secretory growth factors often associated with extracellular matrix (ECM) and extracellular vesicles (EVs) such as exosomes or matrix-coated vesicles. CCN factors and fragments loaded on/in EVs may play key roles in cell communication networks in cancer biology, bone and cartilage metabolism, wound healing, and tissue regeneration. CCN proteins and EVs/exosomes are found in body fluids, such as blood, urine, milk, and supernatants of the two-dimensionally (2D) cultured cells and three-dimensionally (3D) cultured tissues, such as spheroids or organoids. More than ten methods to isolate exosomes or EVs have been developed with different properties. Here, we introduce comprehensive protocols for polymer-based precipitation, affinity purification, ultracentrifugation methods combined with the ultrafiltration method for isolating CCN-loaded exosomes/EVs from 2D and 3D cultured tissues, and proteome analysis using mass spectrometry for comprehensive analysis of CCN proteins.",
keywords = "CCN factors, Exosome isolation methods, Extracellular vesicles, Mass spectrometry, Polymer-based precipitation, Proteome analysis, Spheroid, Tumoroid, Ultracentrifugation, Ultrafiltration",
author = "Takanori Eguchi and Yuka Okusya and Yanyin Lu and Kisho Ono and Taha, {Eman A.} and Shiro Fukuoka",
note = "Funding Information: T.E. was supported by JSPS Kakenhi, grant numbers 17K11642-TE, 20K09904-CS, 19H03817-MT, 20H03888-HN, 20K20611-MT, 20H03888-HN, 21H03119-TY, and 21K08902-HY. Y.O. was supported by JSPS Overseas Research Fellowship. K.O. was supported by JSPS Kakenhi, grant number 21K17115. S.F. was supported by JSPS Kakenhi, grant number 21K20992. Y.L was supported by Rotary Yoneyama Memorial Foundation Fellowship. E.A.T. was supported by the Egypt-Japan Education Partnership Grant. The authors thank Heeyoun Bunch, Junya Futagawa, Kazuko Kobayashi, Kuniaki Okamoto, Chiharu Sogawa, Masaharu Takigawa, Masaki Matsumoto, Seiji Tamaru, Koji Ueda, and Haruo Urata for useful information, discussion, materials, or experimentation. Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2023",
doi = "10.1007/978-1-0716-2744-0_6",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "59--76",
booktitle = "Methods in Molecular Biology",
}
