Trehalose transporter from African chironomid larvae improves desiccation tolerance of Chinese hamster ovary cells

TY - JOUR

T1 - Trehalose transporter from African chironomid larvae improves desiccation tolerance of Chinese hamster ovary cells

AU - Chakraborty, Nilay

AU - Menze, Michael A.

AU - Elmoazzen, Heidi

AU - Vu, Halong

AU - Yarmush, Martin L.

AU - Hand, Steven C.

AU - Toner, Mehmet

N1 - Funding Information: This work was supported by National Institutes of Health Grants 2 RO1 DK046270-14A1 and 5 R01 GM086886. We thank Dr. Takahiro Kikawada at National Institute of Agrobiological Sciences, Ibaraki, Japan for providing us with the trehalose transporter. We thank Dr. Alireza Abazari for helpful discussions.

PY - 2012/4

Y1 - 2012/4

N2 - Dry preservation has been explored as an energy-efficient alternative to cryopreservation, but the high sensitivity of mammalian cells to desiccation stress has been one of the major hurdles in storing cells in the desiccated state. An important strategy to reduce desiccation sensitivity involves use of the disaccharide trehalose. Trehalose is known to improve desiccation tolerance in mammalian cells when present on both sides of the cell membrane. Because trehalose is membrane impermeant the development of desiccation strategies involving this promising sugar is hindered. We explored the potential of using a high-capacity trehalose transporter (TRET1) from the African chironomid Polypedilum vanderplanki [21] to introduce trehalose into the cytoplasm of mammalian cells and thereby increase desiccation tolerance. When Chinese hamster ovary cells (CHO) were stably transfected with TRET1 (CHO-TRET1 cells) and incubated with 0.4 M trehalose for 4 h at 37 °C, a sevenfold increase in trehalose uptake was observed compared to the wild-type CHO cells. Following trehalose loading, desiccation tolerance was investigated by evaporative drying of cells at 14% relative humidity. After desiccation to 2.60. g of water per gram dry weight, a 170% increase in viability and a 400% increase in growth (after 7 days) was observed for CHO-TRET1 relative to control CHO cells. Our results demonstrate the beneficial effect of intracellular trehalose for imparting tolerance to partial desiccation.

AB - Dry preservation has been explored as an energy-efficient alternative to cryopreservation, but the high sensitivity of mammalian cells to desiccation stress has been one of the major hurdles in storing cells in the desiccated state. An important strategy to reduce desiccation sensitivity involves use of the disaccharide trehalose. Trehalose is known to improve desiccation tolerance in mammalian cells when present on both sides of the cell membrane. Because trehalose is membrane impermeant the development of desiccation strategies involving this promising sugar is hindered. We explored the potential of using a high-capacity trehalose transporter (TRET1) from the African chironomid Polypedilum vanderplanki [21] to introduce trehalose into the cytoplasm of mammalian cells and thereby increase desiccation tolerance. When Chinese hamster ovary cells (CHO) were stably transfected with TRET1 (CHO-TRET1 cells) and incubated with 0.4 M trehalose for 4 h at 37 °C, a sevenfold increase in trehalose uptake was observed compared to the wild-type CHO cells. Following trehalose loading, desiccation tolerance was investigated by evaporative drying of cells at 14% relative humidity. After desiccation to 2.60. g of water per gram dry weight, a 170% increase in viability and a 400% increase in growth (after 7 days) was observed for CHO-TRET1 relative to control CHO cells. Our results demonstrate the beneficial effect of intracellular trehalose for imparting tolerance to partial desiccation.

KW - Anhydrobiosis

KW - Desiccation tolerance

KW - Lyopreservation

KW - Trehalose

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U2 - 10.1016/j.cryobiol.2011.11.007

DO - 10.1016/j.cryobiol.2011.11.007

M3 - Article

C2 - 22155480

SN - 0011-2240

VL - 64

SP - 91

EP - 96

JO - Cryobiology

JF - Cryobiology

IS - 2

ER -