Exposure of Seminiferous Tubules of Immature Rats to Cryoprotective Media of Various Compositions
Keywords:seminiferous tubules, immature rats, dimethyl sulfoxide, glycerol, polyethylene oxide, bovine serum albumin
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
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
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
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
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.
Methods of clinical and experimental research in medicine / Ed. I.P. Kaidasheva. Poltava: Polimet; 2003.
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.А., Yukhta M.S., Yurchuk Т.А. 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
How to Cite
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).