Cryoprotective Efficiency of Media Combining Oxyethylated Methylcellosolve and Dimethyl Acetamide during Freeze-Thawing of Human Erythrocytes

Aleksandra V. Nikolenko, Olga V. Vyazovska, Valentina V. Chekanova


Using the media, which combine cryoprotectants being chemicals of different classes, is prospective when developing cryoprotective solutions to cryopreserve erythrocytes. This investigation was performed to evaluate cryoprotective activity of media combining representative of polyols, non-penetrating cryoprotectant oxyethylated methyl cellosolve with polymerization degree of n = 33–35 (OEMCn=33–35) and the representative of amides, penetrating cryoprotectant dimethyl acetamide (DMAc). Combinations of OEMCn=33–35 and DMAc in concentration (w/w) ratios 1:1 (10 and 10%); 2:1 (10 and 5%); 3:1 (15 and 5%); 4:1 (20 and 5%); 5:1 (25 and 5%) were compared with 20 and 30% OEMCn=33–35 solutions. Addition of DMAc to cryoprotective media based on OEMCn=33–35 increased the post-thaw osmotic resistance of erythrocytes. Application of the media combining OEMCn=33–35 and DMAc in ratios of 1:1 (10 and 10%) and 3:1 (15 and 5%) provided a high post-thaw survival of erythrocytes. Osmotic fragility of erythrocytes was found to correlate with the indices of intracellular content of potassium and sodium and with hematocrit value during exposure of the cells in cryoprotective media.

Probl Cryobiol Cryomed 2013; 23(4):297–308.


erythrocytes; cryopreservation; multicomponent cryoprotective media; oxyethylated methyl cellosolve; dimethyl acetamide

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Babijchuk L.A., Zemlyanskikh N.G. Optimization and advantages of washing-out method for erythrocytes cryopreservation with PEO-1500. Problems of Cryobiology 2001; (1): 35–41.

Belous A.M., Grischenko V.I. Cryobiology. Kiev: Naukova Dumka, 1994.

Bidnyy S.Yu, Averyanov M.V. Serebryakov V.G. et al. Phase transformations in dimethyl acetamide water system. In: Physicochemical properties and biological effect of cryoprotectants: Collection of scientific papers. Kharkov, 1990. – P. 5–8.

Bogdanchikova O.A., Kireev V.A., Khodko A.T., Kompaniets A.M. Platelet cryopreservation. 2. Efficiency of cryopreservatives based on cryoprotectant combinations under different freezing regimens. Problems of Cryobiology 2010; 20(4): 443–451.

Gulevsky A.K., Bondarenko V.A., Belous A.M. Barrier properties of biomembranes at low temperatures. Kiev: Naukova Dumka, 1988.

Zinchenko A.V., Krasnikova A.O., Musatova I.B. et al. Phase behaviour of hydrooxyethylated methyl cellosolve system with polymerization degree of n=33–35 below 273 K. In: Proceeding of the 5th Ukrainian Biophysical Society Meeting. – Lutsk, 2011. – P. 152.

Kompaniets A.M., Nikolenko A.V., Chekanova V.V., Trots Yu.P. Cryopreservation of erythrocytes under oligomer of oxyethylated glycerol (n = 25). Problems of Cryobiology 2005; 15 (3): 561–565.

Linnik T.P. Amides of aliphatic acids are effective cryoprotectants. II. Cryoprotective properties of compounds of amides series. Problems of Cryobiology 1999; (2): 22–31.

Linnik T.P., Martynyuk I.N., Gaviley O.V., Beletsky E.M. Cytotoxic effect of diols, amides and their mixtures on fowl and turkey sperm prior to freezing. Problems of Cryobiology 2009; 19 (4): 383–394.

Loevsky M.M., Vorotilin A.M., Gulevsky A.K., Belous A.M. Dynamics of content of cations and phosphoorganic compounds in erythrocytes after low-temperature preservation (–196°C) under protection of 1,2-propane diol and glycerol. Bull Eksper Biol 1982; 94 (9): 95–97.

Menshikov V.V. Laboratory methods of research in clinic: Methods of hematological studies. Moscow: Meditsyna, 1987.

Nikolenko A.V., Vyazovskaya O.V. Studying survival of erythro-cytes depending on composition of cryoprotective media based on non-penetrating cryoprotectant oxyethylated methyl cellosolve. Visnyk Kharkivskogo Natsionalnogo Universytetu Imeni V.N. Karazina. Series: Biologiya 2011; 947 (13): 152–158.

Novikov A.N., Kuleshova L.G., Linnik T.P. Mechanisms of growth of ice crystals in complex biology systems. Biofizika 1991; 36 (1): 122–127.

Pushkar N.S., Belous A.M. Introduction into cryobiology. Kiev: Naukova Dumka, 1975.

Ramazanov V.V. Cryoprotective efficiency of medium combining non-penetrating and penetrating cryoprotectants when freezing erythrocyte suspensions of various volumes. Problems of Cryobiology and Cryomedicine 2013; 23 (2): 124–134.

Pat.1622995 of USSR, IPC A61N1/02. Cryoprotectant for human erythrocytes / L.P. Bredikhina, O.V. Lipina, L.A. Khanina et al.; N4489413/30-14; Filed 03.10.1988; Publ. 1991; Bull. 1.

Pat. 2326532 of Russian Federation, IPC A01N1/02. Cryopreservation method of platelets / A.A. Kostyaev, F.S. Sherstnev, S.V. Utemov et al.; Filed 29.08.2006; Publ. 20.06.2008; Bull. 17.

Crawford A., Harris H. Balancing act: Na+ sodium K+ potassium. Nursing 2011; 41 (7): 44–50.

Edwards S. Regulation of water, sodium and potassium: implications for practice. Nursing Standard 2001; 15(22): 36–42.

Glafke C., Akhoondi M., Oldenhof H. et al. Cryopreservation of platelets using trehalose: the role of membrane phase behavior during freezing. Biotechnol Prog 2012; 28 (5): 1347–1354.

Growe H., Oliver. A.E., Yoekstra F.A., Growe L.M. Stabilization of dry membrane by mixtures of hydroxyethyl starch and glucose: the role of vitrification. Cryobiology 1997; 35 (1): 20–30.

Nikolenko A.V., Chekanova V.V., Schetinsky M.I., Vyazovska O.V. Relation between cryoprotective and physico-chemical properties of oxyethylated methyl cellosolve-based media. Cryoletters 2013; 34 (5): 527–534.

Petrenko Y.A., Jones D.R.E., Petrenko A.Y. Cryopreservation of human fetal liver hematopoietic stem / progenitor cells using sucrose as an additive to the cryoprotective medium. Cryobiology 2008; 57 (3): 195–200.

Quan G.B., Han Y., Liu M.X., Gao F. Effects of pre-freeze incubation of human red blood cells with various sugars on postthaw recovery when using a dextran-rapid cooling protocol. Cryobiology 2009; 59 (3): 258–267.

Quan G.B., Han Y., Liu M.X. et al. Addition of oligosaccharide decreases the freezing lesions on human red blood cell membrane in the presence of dextran and glucose. Cryobiology 2011; 62 (2): 135–144.

Savitz D., Sidel V.W., Solomon A.K. Osmotic properties of human red cells. J Gen Physiol 1964; 48(1): 79–94.

Valeri C.R. Blood banking and the use of frozen blood products. – Cleveland: CRC Press, 1976.

Zhivotova E.N., Zinchenko A.V., Kuleshova L.G., Chekanova V.V., Kompaniets A.M. Physical states of aqueous solutions of oxyethylated glycerol with polymerization degree of n = 30 at temperatures lower than 283 K. CryoLetters 2007; 28 (4): 261–270.


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