Trehalose uptake and dehydration effects on the cryoprotection of CHO-K1 cells expressing TRET1

Uchida, Tsutomu; Furukawa, Maho; Kikawada, Takahiro; Yamazaki, Kenji; Gohara, Kazutoshi

Trehalose uptake and dehydration effects on the cryoprotection of CHO-K1 cells expressing TRET1

Uchida, Tsutomu; Furukawa, Maho; Kikawada, Takahiro et al.

Cryobiology, 2019, 90, 30-40

Permalink : https://hdl.handle.net/2115/79357

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論文情報

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アクセス権	open access
URI	https://hdl.handle.net/2115/79357
タイトル	Trehalose uptake and dehydration effects on the cryoprotection of CHO-K1 cells expressing TRET1
著者	Uchida, Tsutomu ; Furukawa, Maho ; Kikawada, Takahiro ; Yamazaki, Kenji ; Gohara, Kazutoshi
e-Rad_Researcher 70356575 著者名 Uchida, Tsutomu
著者名 Furukawa, Maho
著者名 Kikawada, Takahiro
著者名 Yamazaki, Kenji
e-Rad_Researcher 40153746 著者名 Gohara, Kazutoshi
言語	英語
キーワード	Trehalose transporter
	Freeze thaw viability
	Immersion time
	Maximum uptake
	Apoptosis rate
	Optimum conditions
発行日	2019-10
出版者	Elsevier
収録物名	Cryobiology
巻(号)	90
ページ	30-40
抄録	Chinese hamster ovary cells (CHO K1 cells) in which the trehalose transporter (TRET1) is expressed can have greater cryoprotection than ordinary CHO Ki cells. This study examines the uptake characteristics of trehalose into cells via TRET1 and determines the influence of intracellular trehalose on the freeze thaw viabilities. In our experiments, the intracellular trehalose concentration is controlled by the extracellular trehalose concentration and the immersion time in a freezing solution. In this freezing solution, both kinds of CHO Kl cells are independently dispersed with various amount of trehalose, and then put into the CO2 incubator for 0-6 h. After a set immersion time, the cell -suspended sample is cooled to 193 K, stored for 1 week, then quickly thawed at 310 K and its viability measured. The uptake amount of intracellular trehalose is measured before freezing. We find an upper limit for the uptake amount of trehalose when the extracellular trehalose concentration is about 400 mM, at which the freeze thaw viability is the highest. When the extracellular trehalose concentration exceeds 400 mM, shorter immersion times are needed to obtain the maximum freeze thaw viability. Also, longer immersion weakens the cells. Our analyses indicate that when the extracellular trehalose-concentration is less than 400 mM, the trehalose uptake occurs more slowly with less dehydration, resulting in less stress on the cell. When the extracellular trehalose concentration exceeds the saturation level, the cell is stressed by the excess dehydration due to the remaining osmotic pressure, with apoptosis occurring before freezing.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
権利情報	© 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
PISSN	0011-2240
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.cryobiol.2019.09.002