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

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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 cells and the wild-type cells under various trehalose concentrations in an extracellular medium and under various freezing temperatures. We show here that the intracellular trehalose significantly improves the viability of freeze-thawing cells. The optimum trehalose concentration is about 400 mM, and the optimum freezing temperature ranges between 140 and 240 K. We find a qualitative coincidence of the optimum freezing temperature with that from including 10 vol% dimethyl sulfoxide, a result that indicates that both intra- and extracellular trehalose principally work to inhibit ice crystal growth and thus protect the membrane and proteins from ice crystals during relatively slow freezing.

source:SC30201804090007

source:NARO成果DBa

source:Green open Access

source:This journal has an embargo period of 12 months.

source:この論文は出版社版ではありません。引用の際には出版社版をご利用ください。

source:This is not the published version. Please cite only the published version.

source:https://doi.org/10.1016/j.cryobiol.2017.05.008

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