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Intracellular trehalose via transporter TRET1 as a method to cryoprotect CHO-K1 cells

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Title: Intracellular trehalose via transporter TRET1 as a method to cryoprotect CHO-K1 cells
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
Trehalose transporter 1
Cryoprotectant
Freezing rate
Trehalose concentration
CHO-K1 cell
Issue Date: Aug-2017
Publisher: Elsevier
Journal Title: Cryobiology
Volume: 77
Start Page: 50
End Page: 57
Publisher DOI: 10.1016/j.cryobiol.2017.05.008
PMID: 28552273
Abstract: Trehalose is a promising natural cryoprotectant, but its cryoprotective effect is limited due to difficulties in transmembrane transport. Thus, expressing the trehalose transporter TRET1 on various mammalian cells may yield more trehalose applications. In this study, we ran comparative cryopreservation experiments between the TRET1-expressing CHO-K1 cells (CHO-TRET1) and the CHO-K1 cells transfected with an empty vector (CHO-vector). The experiments involve freezing under various trehalose concentrations in an extracellular medium. The freeze-thawing viabilities of CHO-TRET1 cells are higher than those of CHO-vector cells for most freezing conditions. This result differs from control experiments with a transmembrane type cryoprotectant, dimethyl sulfoxide (Me2SO), which had similar viabilities in each condition for both cell types. We conclude that the trehalose loaded into the cells with TRET1 significantly improves the cryoprotective effect. The higher viabilities occurred when the extracellular trehalose concentration exceeded 200 mM, with 250-500 mM being optimal, and a cooling rate below 30 K/min, with 5-20 K/min being optimal.
Rights: © 2017. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
http://creativecommons.org/licenses/by-nc-nd/4.0/
Type: article (author version)
URI: http://hdl.handle.net/2115/71129
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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