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

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Title: Trehalose uptake and dehydration effects on the cryoprotection of CHO-K1 cells expressing TRET1
Authors: Uchida, Tsutomu Browse this author →KAKEN DB
Furukawa, Maho Browse this author
Kikawada, Takahiro Browse this author
Yamazaki, Kenji Browse this author
Gohara, Kazutoshi Browse this author →KAKEN DB
Keywords: Trehalose transporter
Freeze thaw viability
Immersion time
Maximum uptake
Apoptosis rate
Optimum conditions
Issue Date: Oct-2019
Publisher: Elsevier
Journal Title: Cryobiology
Volume: 90
Start Page: 30
End Page: 40
Publisher DOI: 10.1016/j.cryobiol.2019.09.002
Abstract: 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.
Rights: © 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license
Type: article (author version)
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 内田 努

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