Exposure of Seminiferous Tubules of Immature Rats to Cryoprotective Media of Various Compositions

Authors

  • Natalya O. Volkova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Mariya S. Yukhta Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Lyudmyla G. Chernyshenko Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Lyudmyla V. Stepanyuk Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Larysa V. Sokol Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Anatoliy M. Goltsev Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

DOI:

https://doi.org/10.15407/cryo27.03.203

Keywords:

seminiferous tubules, immature rats, dimethyl sulfoxide, glycerol, polyethylene oxide, bovine serum albumin

Abstract

The development of new methods aimed at preserving potential fertility before the cytotoxic therapy for individuals of pre-adulthood is an actual area of medical and biological research. The research compares the effect of exposure to media of different composition on morphological parameters and metabolic activity of cells of testicular convoluted tubules of sexually immature rats. Cryopreservation of testicular tissues requires the use of appropriate protocols, ensuring the high viability and functional activity at the background of minimization of damages in transplant material. It has been established that a 30-minute exposure of the tissue samples in media based on Hanks solution and 50 g/l bovine serum albumin supplemented with either 0.6 M DMSO or 0.7 M glycerol did not result in the damage of spermatogenic epithelium and to the decrease in metabolic activity of the cells (MTT test and lactate dehydrogenase activity). Exposure of the testicular convoluted tubules of immature rats during 45 and 60 min in cryoprotective media supplemented with 0.6 M DMSO, 0.7 M glycerol, 50 g/L polyethylene oxide with molecular weight 400 and 0.1 M sucrose led to the manifested desquamation, appearance of acellular areas, reduced density of spermatogenic epithelium cells and their metabolic activity. The findings could be used to substantiate and develop the effective techniques for cryopreservation of seminiferous tubules of immature rats.

Probl Cryobiol Cryomed 2017; 27(3): 203–218

Author Biographies

Natalya O. Volkova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryopathophysiology and Immunology

Mariya S. Yukhta, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryopathophysiology and Immunology

Lyudmyla G. Chernyshenko, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryopathophysiology and Immunology

Lyudmyla V. Stepanyuk, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryopathophysiology and Immunology

Larysa V. Sokol, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryopathophysiology and Immunology

Anatoliy M. Goltsev, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryopathophysiology and Immunology

References

Aliakbari F., Yazdekhasti H., Abbasi M. et al. Advances in cryopreservation of spermatogonial stem cells and restoration of male fertility. Microsc Res Tech 2016; 79(2): 122–129. CrossRef PubMed

Avtandilov G.G. Medical morphometry: Manual. Moskow: Medicine; 1990.

Baert Y., Braye A., Struijk R.B. et al. Cryopreservation of testicular tissue before long-term testicular cell culture does not alter in vitro cell. Fertil Steril 2015; 104(5): 1244–1252. CrossRef PubMed

Baust J.M., Vogel M.J., Van Buskirk R., Baust J.G. A molecular basis of cryopreservation failure and its modulation to improve cell survival. Cell Transplant 2001; 10(7): 561–571. CrossRef

Bystrova O.V., Kalugina A.S., Tsybatova E.V. et al. Methods for recovering fertility in cancer patients. Prakt Onkologiya 2009; 10(4): 245–253.

Day J.G., Stacey G.N. Cryopreservation and freeze-drying protocols. – Totowa, New Jersey: Humanna Press; 2007. CrossRef

Dooley D.C., Law P., Schork P., Meryman H.T. Glycerolization of the human neutrophil for cryopreservation: osmotic response of the cell. Exp Hematol 1982; 10: 423–434. PubMed

Easley C., Simerly C., Schatten G. Stem cell therapeutic possibilities: future therapeutic options for male-factor and female-factor infertility? Reprod Biomed Online 2013; 27(1): 75–80. CrossRef PubMed

Fleming K.K., Hubel A. Cryopreservation of hematopoietic and non-hematopoietic stem cells. Transfus Apher Sci 2006; 34(3): 309–315. CrossRef PubMed

Ginsburg E.S., Yanushpolsky E.H., Jackson K.V. In vitro fertilization for cancer patients and survivors. Fertility and Sterility 2001; 75(4): 705–710. CrossRef

Izadyar F., Matthijs-Rijsenbilt J.J., den Ouden K. et al. Development of a cryopreservation protocol for type A spermatogonia. J Androl 2002; 23(4): 537–545. PubMed

Izadyar F., Spierenberg G.T., Creemers L.B. et al. Isolation and purification of type A spermatogonia from the bovine testis. Reproduction 2002; 124(1): 85–94. CrossRef PubMed

Keros V., Hultenby K., Borgstrom B. et al. Methods of cryopreservation of testicular tissue with viable spermatogonia in prepubertal boys undergoing gonadotoxic cancer treatment. Hum Reprod 2007; 22(5): 1384–1395. CrossRef PubMed

Keros V., Rosenlund B., Hultenby K. et al. Optimizing cryopreservation of human testicular tissue: comparison of protocols with glycerol, propanediol and dimethylsulphoxide as cryoprotectants. Hum Reprod 2005; 20(6): 1676–1687. CrossRef PubMed

Kim K.J., Lee Y.A., Kim B.J. et al. Cryopreservation of putative pre-pubertal bovine spermatogonial stem cells by slow freezing. Cryobiology 2015; 70(2): 175–183. CrossRef PubMed

Kuleshova L.G., Gordienko E.A., Kovalenko I.F. Permeability of plasma membranes of rat hepatocytes for water molecules in a non-isotonic medium of electrolytes. Biofizitchny Visnik 2002; 11(2): 39–42.

Lee Y.A., Kim Y.H., Ha S.J. et al. Cryopreservation of porcine spermatogonial stem cells by slow-freezing testis tissue in trehalose1. Journal of Animal Science 2014; 92(3): 984–995. CrossRef PubMed

Methods of clinical and experimental research in medicine / Ed. I.P. Kaidasheva. Poltava: Polimet; 2003.

Moss A.C., Higgins A.Z. Investigating the potential for cryopreservation of human granulocytes with concentrated glycerol. Cryobiology 2016; 72(3): 290–293. CrossRef PubMed

Mossman T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65(1–2): 55–63. CrossRef

Onofre J., Baert Y., Faes K., Goossens E. Cryopreservation of testicular tissue or testicular cell suspensions: a pivotal step in fertility preservation. Human Reproduction Update 2016; 22(6): 744–761. CrossRef PubMed

Pelufo V., Lopez Armengol M.F., Malcotti V. et al. Effects of glycerol and sugar mixing temperature on the morphologic and functional integrity of cryopreserved ram sperm. Theriogenology 2015; 83(1): 144–151. CrossRef PubMed

Poirot C., Schubert B. Fertility preservation in prepubertal children. Bull Cancer 2011; 98(5): 489–499. PubMed

Pushkar N.S., Shrago M.I., Belous A.M., Kalugin Yu.V. Cryoprotectors. Kyiv: Naukova Dumka; 1978.

Rajotte R.V. Islet cryopreservation protocols. Annals of the New York Academy of Sciences 1999; 875(1): 200–207. CrossRef

Smolyaninova E.I., Khromenkova O.B., Zhernoklev G.V., Pishko O.V. Effect of equilibration in the freezing environment on osmotic stability and viability of mouse 8-cell embryos. Probl Cryobiol 2004; 1: 3–11.

Unni S., Kasiviswanathan S., D'Souza S. et al. Efficient cryopreservation of testicular tissue: effect of age, sample state, and concentration of cryoprotectant. Fertil Steril 2012; 97: 200-8.e201. CrossRef PubMed

Volkova N.A., Pavlovich E.V., Gapon A.A., Nikolov O.T. Effects of millimeter-wave electromagnetic exposure on the morphology and function of human cryopreserved spermatozoa. Bulletin of Experimental Biology and Medicine 2014; 157(5): 574–576. CrossRef PubMed

Volkova N.Ð., Yukhta M.S., Yurchuk Т.Ð. et al. Multipotent mesenchymal stromal cells of bone marrow in therapy of chronic inflammation of the murine ovaries. Biotechnologia Acta 2014; 7(5): 35–42. CrossRef

Zhang J.M., Li L.X., Yang Y.X. et al. Is caspase inhibition a valid therapeutic strategy in cryopreservation of ovarian tissue? J Assist Reprod 2009; 26(7): 415–420. CrossRef PubMed

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Published

2017-09-25

How to Cite

Volkova, N. O., Yukhta, M. S., Chernyshenko, L. G., Stepanyuk, L. V., Sokol, L. V., & Goltsev, A. M. (2017). Exposure of Seminiferous Tubules of Immature Rats to Cryoprotective Media of Various Compositions. Problems of Cryobiology and Cryomedicine, 27(3), 203–218. https://doi.org/10.15407/cryo27.03.203

Issue

Section

Theoretical and Experimental Cryobiology