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

Abstract：

: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 0.4M trehalose for 4h 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 2.60g of water per gram dry weight, a 170% increase in viability and a 400% increase in growth (after 7days) 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.

journal_name

Cryobiology

journal_title

Cryobiology

authors

Chakraborty N,Menze MA,Elmoazzen H,Vu H,Yarmush ML,Hand SC,Toner M

doi

10.1016/j.cryobiol.2011.11.007

subject

Has Abstract

pub_date

2012-04-01 00:00:00

pages

91-6

issue

2

eissn

0011-2240

issn

1090-2392

pii

S0011-2240(11)00328-2

journal_volume

64

pub_type

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