Cryoprotective Efficiency of Medium Combining Non-Penetrating and Penetrating Cryoprotectants When Freezing Erythrocyte Suspensions of Various Volumes

Viktor V. Ramazanov

Abstract


Osmotic properties of erythrocytes, frozen in suspesions with different volumes in combined medium with 15% dextran and 5% DMSO were studied. It was established that the increasing of sample volume subjected to freezing and reduction of freezing and thawing rate did not result in a significant rise of H+ ion flow and loss of membrane barrier function in respect of glutathione in cells washed free of cryopreservative. The obtained results allowed to suggest that freezing in combined medium independent on cooling rate provided optimal dehydration of cells. It was determined that inclusion of DMSO into a cryopreservative based on dextran and its penetration into cells decreased dehydration and hypertonic stress rate, stipulated by concentrating of non-penetrating components of medium during freezing. Lowering of hypertonic stress was a pre-condition of erythrocyte membrane resistance to post-hypertonic stress during freeze-thawing and contributed to preservation of osmotic properties of cells washed free of cryopreservative.


Probl Cryobiol Cryomed 2013; 23(2):124–134.


Keywords


combined cryopreservatives; dextran; dimethyl sulfoxide; erythrocytes; osmotic properties; membrane barrier properties; glutathione; sulfate

Full Text:

PDF (RU+EN)

References


Ashmarin I.P., Vasil'ev I.P., Ambrosov V.A. Express methods of statistical analysis and planning of experiments. Leningrad, 1975.

Mezhidov S.Kh., Belyaeva I.M., Vorotilin A.M., Moiseev V.A. Effect of combination of polyvinylpirrolidon and 1,2-propane-diol to the erythrocytes survival at cryopreservation. Problems of Cryobiology 1996; (2): 22–25.

Mezhidov S.Kh., Moiseev V.A. Effect of combination of high molecular cryoprotectants with 1,2–propanediol to the erythrocytes survival at cryopreservation. Problems of Cryobiology 1995; (3): 46–48.

Ramazanov V.V. Effect of combined media to erythrocytes destruction frozen with different hematocrit. Problems of Cryobiology 2006; 16 (2): 155–163.

Ramazanov V.V., Zabrodsky R.F., Naydyuk Ya.Yu., Bondarenko V.A. Functioning of H+ ion transport system during modification of erythrocyte membranes under conditions, modeling freezing ones. Visnyk Problem Biologii i Medytsyny 2010; (3): 186–192.

Sakun O.V. Analysis of cryopreservation efficiency of cell suspensions with constant and changed cooling rate [Thesis of Candidate of Biol. Sciences]. Kharkiv, 2009.

Reference book for physical and technological principals of genetics. Ed. by M.P. Malkova. Moscow: Energiya, 1973.

Bakhach J. The cryopreservation of composite tissues. Principles and resent advancement on cryopreservation of different type of tissues. Organogenesis 2009; 5 (3): 119–126.

Beutler E. Red cell metabolism. A manual of biochemical methods. New York: Grune&Stratton, 1975.

Brahm J. Temperature-dependent changes of chloride transport kinetics in human red cells. J Gen Physiol 1977; 70 (3): 283–306.

Clapisson G., Salinas С., Malacher P. et al. Cryopreservation with hydroxyethylstarch (HES) + dimethylsulphoxide (DMSO) gives better results than DMSO alone. Bull Cancer 2004; 91 (4): E97–102.

Devireddy R.V., Swanlund D.J., Olin T. et al. Cryopreservation of equine sperm: optimal cooling rates in the presence and absence of cryoprotective agents determined using differential scanning calorimetry. Biol Reprod 2002; 66 (1): 222–231.

Farrant J., Walter C.A., Lee H., McGann L.E. Use of two-step cooling procedures to examine factors influencing cell survival following freezing and thawing. Cryobiology 1977; 14 (3): 273–286.

Levin R.L. The limiting effects of heat and mass transfer on the osmotic behavior of cells during freezing and thawing. Cryobiology 1982; 19 (6): 669.

Lionetti F.J., Luscinskas F.W., Hant S.M. et al. Factors affecting the stability of cryogenically preserved granulocytes. Cryobiology 1980; 17 (3): 297–310.

Liu K.Y., Dong W.C., Wang Y.L. et al. Study on non-programmed process using dimethyl sulfoxide and hydroxyethyl starch as cryoprotectants in cryopreservation of cord blood hematopoietic cells. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2004; 12 (5): 670–673.

Luo K., Wu G., Wang Q. et al. Effect of dimethylsulfoxide and hydroxyethyl starch in the preservation of fractionated human marrow cells. Cryobiology 1994; 31 (4): 349–354.

Maruyama M., Kenmochi T., Sakamoto K. et al. Simplified method for cryopreservation of islets using hydroxyethyl starch and dimethyl sulfoxide as cryoprotectants. Transplant. Proc 2004; 36 (4): 1133–1134.

Mazur P. The role of intracellular freezing in the death of cells cooled at supraoptimal rates. Cryobiology 1977; 14 (3): 251–272.

Pegg D.E., Diaper M.P. The paсking effect in erythrocyte freezing. Cryo-Letters 1983; 4 (2): 129–136.

Pitt R.E., Chandrasekaran M., Parks J.E. Performance of a kinetic model for intracellular ice formation based on the extent of supercooling. Cryobiology1992; 29 (3): 359–373.

Reiners B., Zintl F., Plenert W. Use of human albumin as an additional cryoprotective agent in freeze preservation of hematopoietic stem cells. Folia Haematol Int Mag Klin Morphol Blutforsch 1987; 114 (2): 264–272.

Romano L., Passow H. Characterization of anion transport system in trout red blood cell. Am J Physiol 1984; 246: 330–338.

Rowley S.D., Feng Z., Chen L. et al. A randomized phase III clinical trial of autologous blood stem cell transplantation comparing cryopreservation using dimethylsulfoxide vs dimethylsulfoxide with hydroxyethyl starch. Bone Marrow Transplant 2003; 31 (11): 1043–1051.

Stiff P.J., Koester A.R., Weidner M.K. et al. Autologous bone marrow transplantation using unfractionated cells cryopreserved in dimethylsulfoxide and hydroxyethyl starch without controlled-rate freezing. Blood 1987; 70 (4): 974–978.

Stiff P.J., Murgo A.J., Zaroulis C.G. et al. Unfractionated human marrow cell cryopreservation using dimethylsulfoxide and hydroxyethyl starch. Cryobiology 1983; 20 (1): 17–24.

Tsuruta T., Ishimoto Y., Masuoka T. Effects of glycerol on intracellular ice formation and dehydration of onion epidermis. Ann NY Acad Sci 1998; 858: 217–226.

Wagner C.T., Martowicz M.L., Livesey S.A., Connor J. Biochemical stabilization enhances red blood cell recovery and stability following cryopreservation. Cryobiology 2002; 45 (2): 153–166.

Woelders H., Chaveiro A. Theoretical prediction of 'optimal' freezing programmes. Cryobiology 2004; 49 (3): 258–271.


Refbacks

  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

 

Institute for Problems of Cryobiology and Cryomedicine

23, Pereyaslavskaya str., Kharkov, Ukraine

Tel. +38057 373 4143; Fax +38057 373 5952

e-mail: journal@cryo.org.ua