Introduction of Lyo- and Cryoprotective Carbohydrates into Mammalian Cells through Swelling-Activated Channels

Abstract

As sugars and related compounds are increasingly being used as potential cryo- and lyoprotective agents for preservation of rare and valuable mammalian cells and tissues, introduction of these compounds, especially saccharide, into a living cell is one of the key technology. In the present study, volume changes of human T-lymphocytes (Jurkat line) exposed to hypotonic carbohydrate-substituted solutions of different composition and osmolality were studied by videomicroscopy. The complex volumetric data were analyzed with a membrane transport model that allowed the estimation of the hydraulic conductivity and volume-dependent solute permeabilities. We found that under hypotonic stress of 100 mOsm, the membrane permeability to monomeric carbohydrates inereased dramatically (apparently due to channel activation caused by extensive cell swelling), whereas oligosaccharide permeability remained very poor. The size-selectivity of the swelling-activated sugar permeation was confirmed by direct chromatographic measurements of intracellular sugars. The results of this study are of interest for biotechnology.