$Pentapartite fractionation of particles in oral fluids by differential centrifugation$
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Pentapartite fractionation of particles in oral fluids by differential centrifugation

Abstract Oral fluids (OFs) contain small extracellular vesicles (sEVs or exosomes) that carry disease-associated diagnostic molecules. However, cells generate extracellular vesicles (EVs) other than sEVs, so the EV population is quite heterogeneous. Furthermore, molecules not packaged in EVs can als...

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Autores principales:	Chiho Hiraga, Satoshi Yamamoto, Sadamitsu Hashimoto, Masataka Kasahara, Tamiko Minamisawa, Sachiko Matsumura, Akira Katakura, Yasutomo Yajima, Takeshi Nomura, Kiyotaka Shiba
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8a59af34df4a4589b23eb9954ca6dabd
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spelling	oai:doaj.org-article:8a59af34df4a4589b23eb9954ca6dabd2021-12-02T14:11:28ZPentapartite fractionation of particles in oral fluids by differential centrifugation10.1038/s41598-021-82451-62045-2322https://doaj.org/article/8a59af34df4a4589b23eb9954ca6dabd2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82451-6https://doaj.org/toc/2045-2322Abstract Oral fluids (OFs) contain small extracellular vesicles (sEVs or exosomes) that carry disease-associated diagnostic molecules. However, cells generate extracellular vesicles (EVs) other than sEVs, so the EV population is quite heterogeneous. Furthermore, molecules not packaged in EVs can also serve as diagnostic markers. For these reasons, developing a complete picture of particulate matter in the oral cavity is important before focusing on specific subtypes of EVs. Here, we used differential centrifugation to fractionate human OFs from healthy volunteers and patients with oral squamous cell carcinoma into 5 fractions, and we characterized the particles, nucleic acids, and proteins in each fraction. Canonical exosome markers, including CD63, CD9, CD133, and HSP70, were found in all fractions, whereas CD81 and AQP5 were enriched in the 160K fraction, with non-negligible amounts in the 2K fraction. The 2K fraction also contained its characteristic markers that included short derivatives of EGFR and E-cadherin, as well as an autophagosome marker, LC3, and large multi-layered vesicles were observed by electronic microscopy. Most of the DNA and RNA was recovered from the 0.3K and 2K fractions, with some in the 160K fraction. These results can provide guideline information for development of purpose-designed OF-based diagnostic systems.Chiho HiragaSatoshi YamamotoSadamitsu HashimotoMasataka KasaharaTamiko MinamisawaSachiko MatsumuraAkira KatakuraYasutomo YajimaTakeshi NomuraKiyotaka ShibaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Chiho Hiraga Satoshi Yamamoto Sadamitsu Hashimoto Masataka Kasahara Tamiko Minamisawa Sachiko Matsumura Akira Katakura Yasutomo Yajima Takeshi Nomura Kiyotaka Shiba Pentapartite fractionation of particles in oral fluids by differential centrifugation
description	Abstract Oral fluids (OFs) contain small extracellular vesicles (sEVs or exosomes) that carry disease-associated diagnostic molecules. However, cells generate extracellular vesicles (EVs) other than sEVs, so the EV population is quite heterogeneous. Furthermore, molecules not packaged in EVs can also serve as diagnostic markers. For these reasons, developing a complete picture of particulate matter in the oral cavity is important before focusing on specific subtypes of EVs. Here, we used differential centrifugation to fractionate human OFs from healthy volunteers and patients with oral squamous cell carcinoma into 5 fractions, and we characterized the particles, nucleic acids, and proteins in each fraction. Canonical exosome markers, including CD63, CD9, CD133, and HSP70, were found in all fractions, whereas CD81 and AQP5 were enriched in the 160K fraction, with non-negligible amounts in the 2K fraction. The 2K fraction also contained its characteristic markers that included short derivatives of EGFR and E-cadherin, as well as an autophagosome marker, LC3, and large multi-layered vesicles were observed by electronic microscopy. Most of the DNA and RNA was recovered from the 0.3K and 2K fractions, with some in the 160K fraction. These results can provide guideline information for development of purpose-designed OF-based diagnostic systems.
format	article
author	Chiho Hiraga Satoshi Yamamoto Sadamitsu Hashimoto Masataka Kasahara Tamiko Minamisawa Sachiko Matsumura Akira Katakura Yasutomo Yajima Takeshi Nomura Kiyotaka Shiba
author_facet	Chiho Hiraga Satoshi Yamamoto Sadamitsu Hashimoto Masataka Kasahara Tamiko Minamisawa Sachiko Matsumura Akira Katakura Yasutomo Yajima Takeshi Nomura Kiyotaka Shiba
author_sort	Chiho Hiraga
title	Pentapartite fractionation of particles in oral fluids by differential centrifugation
title_short	Pentapartite fractionation of particles in oral fluids by differential centrifugation
title_full	Pentapartite fractionation of particles in oral fluids by differential centrifugation
title_fullStr	Pentapartite fractionation of particles in oral fluids by differential centrifugation
title_full_unstemmed	Pentapartite fractionation of particles in oral fluids by differential centrifugation
title_sort	pentapartite fractionation of particles in oral fluids by differential centrifugation
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8a59af34df4a4589b23eb9954ca6dabd
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Pentapartite fractionation of particles in oral fluids by differential centrifugation

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