Intracellular trehalose via transporter TRET1 as a method to cryoprotect CHO-K1 cells |
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| Affiliation: | 1. Division of Applied Physics, Faculty of Engineering, Hokkaido University, N13 W8 Kita-ku, Sapporo, Hokkaido 060-8628, Japan;2. Anhydrobiosis Research Group, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Ohwashi 1-2, Tsukuba, Ibaraki 305-8634, Japan;3. Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5, Kashiwanoha, Kashiwa, Chiba 277-8561, Japan;1. Laboratory of Cell Biology, Institute of Microbiology and Biotechnology, University of Latvia, Jelgavas Str., 1-537, LV-1004, Riga, Latvia;2. Department of Agricultural, Food and Environmental Science & Industrial Yeasts Collection DBVPG, University of Perugia, Borgo XX Giugno 74, I-06121, Perugia, Italy;3. Department of Genetics, Institute of Molecular and Cell Biology, University of Tartu, Riia 23, 51010, Tartu, Estonia;4. Institute for Global Food Security, School of Biological Sciences, MBC, Queen''s University Belfast, Belfast, BT9 7BL, Northern Ireland, UK;1. Institute of Technology and Science, The University of Tokushima, Minamijosanjima-cho 2-1, Tokushima 770-8506, Japan;2. Advanced Technology and Science, The University of Tokushima, Minamijosanjima-cho 2-1, Tokushima 770-8506, Japan;3. Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, 6-6-11-606 Aoba-yama, Aramaki, Aoba-ku, Sendai 980-8579, Japan;4. Department of Biotechnology, Graduate School of Engineering, Osaka University, Yamadaoka 2-1, Suita, Osaka 565-0871, Japan;1. Department of Chemical Engineering, Faculty of Engineering, Graduate School, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka-city, Fukuoka 819-0395, Japan;2. Global Innovation Center, Kyushu University, Fukuoka Industry-Academia Symphonicity, 4-1, Kyudai-Shinmachi, Nishi-ku, Fukuoka-city, Fukuoka 819-0388, Japan |
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| 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. |
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| Keywords: | Trehalose Trehalose transporter 1 Cryoprotectant Freezing rate Trehalose concentration CHO-K1 cell TRET1" },{" #name" :" keyword" ," $" :{" id" :" kwrd0045" }," $$" :[{" #name" :" text" ," _" :" trehalose transporter 1 dimethyl sulfoxide SD" },{" #name" :" keyword" ," $" :{" id" :" kwrd0065" }," $$" :[{" #name" :" text" ," _" :" standard deviation calcein-AM" },{" #name" :" keyword" ," $" :{" id" :" kwrd0075" }," $$" :[{" #name" :" text" ," _" :" acetoxymethylated calcein PI" },{" #name" :" keyword" ," $" :{" id" :" kwrd0085" }," $$" :[{" #name" :" text" ," _" :" propidium iodide |
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