Intracellular delivery of carbohydrates into mammalian cells through swelling-activated pathways.
Volume changes of human T-lymphocytes (Jurkat line) exposed to hypotonic carbohydrate-substituted solutions of different composition and osmolality were studied by videomicroscopy. In 200 mOsm media the cells first swelled within 1-2 min and then underwent regulatory volume decrease (RVD) to their original isotonic volume within 10-15 min. RVD also occurred in strongly hypotonic 100 mOsm solutions of di- and trisaccharides (trehalose, sucrose, raffinose). In contrast to oligosaccharide media, 100 mOsm solutions of monomeric carbohydrates (glucose, galactose, inositol and sorbitol) inhibited RVD. 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 slightly hypotonic stress (200 mOsm) the cell membrane was impermeable to all carbohydrates studied here. Upon osmolality decrease to 100 mOsm, the membrane permeability to monomeric carbohydrates increased 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, where sugars and related compounds are increasingly being used as potential cryo- and lyoprotective agents for preservation of rare and valuable mammalian cells and tissues.
Reuss
R
R
Lehrstuhl für Biotechnologie, Biozentrum, Universität Würzburg, Am Hubland, Würzburg, Germany.
Ludwig
J
J
Shirakashi
R
R
Ehrhart
F
F
Zimmermann
H
H
Schneider
S
S
Weber
M M
MM
Zimmermann
U
U
Schneider
H
H
Sukhorukov
V L
VL
D001692:Biological Transport; D050260:Carbohydrate Metabolism; D002463:Cell Membrane Permeability; D048429:Cell Size; D003451:Cryoprotective Agents / Q000378:metabolism; D006801:Humans; D007038:Hypotonic Solutions; D019169:Jurkat Cells; D018715:Microscopy, Video
D003451:Cryoprotective Agents; D007038:Hypotonic Solutions