Abstract
Electron paramagnetic resonance of spin probes was used to study the effect of low and high molecular weight CPAs on microviscosity of erythrocyte membranes. Research data on rotational mobility of the fatty acid spin probes in erythrocyte membranes after 5 min incubation in 15 % propylene glycol solution indicate the absence of strong changes in membrane microviscosity. Introduced polyethylene glycol with m. w. 1500 (PEG-1500) at a final concentration of 15% has been shown to increase the membrane microviscosity at 60 min exposure. This is explained by the fact that the molecules of PEG-1500 do not pass inside the cells and are localized on the membrane surface to cause immobilization and inhibiting the rotational mobility of the spin probe. In the initial period of time under these concentrations of PEG-1500 the injury of liquid crystal membrane structure and disorder of the protein-lipid interactions in the membrane prevail over the dehydration of cells. Probl Cryobiol Cryomed 2016; 26(1):35-44.
Highlights
Electron paramagnetic resonance of spin probes was used to study the effect of low and high molecular weight cryoprotective agents (CPAs) on microviscosity of erythrocyte membranes
There is a necessity in investigating the effects of low and high molecular weight CPAs, which are capable of penetrating through the cell cytoplasmic membrane, on microviscosity of membrane lipid phase, and using this knowledge to justify the possibility of application of these CPAs when developing the techniques to cryopreserve the cell suspensions
CPAs propylene glycol (PG) and PEG-1500 were added into the suspension of erythrocytes under the concentration close to those used for cryopreservation of cells
Summary
Electron paramagnetic resonance of spin probes was used to study the effect of low and high molecular weight CPAs on microviscosity of erythrocyte membranes. The following protective mechanisms of the CPAs are among the described in publications: preventing the damage with high saline concentrations in solution [20]; reduction of cell damage, in particular, of cell membranes during dehydration [5, 12, 28]; preventing an intracellular crystallization [10, 25], etc These compounds could render a damaging effect, which extent depends on a medium temperature, concentration of substances and duration of exposure in a solution and can lead to an increased heterogeneity of membrane lipid bilayer, compromised barrier properties, inhibited activity of cytoplasmic and membrane enzyme complexes [1, 5, 12]. The signal from more motile probes appeared in the spectra of electron spin resonance (ESR), that indicated an increased dynamic heterogeneity of erythrocyte membranes due to the emergence of more fluid areas These changes were not found if low molecular weight CPA, propylene glycol, was present in the cell suspension. Optimal incubation time for each cell type is usually experimentally found [2, 16, 27, 28]
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