Importance of fetal bovine serum introduction in hyperosmolar solutions of 1,2-propanediol for preserving morphological integrity of ovarian tissue

  • І. А. Трутаєва
  • В. В. Кірошка
  • А. А. Гавас
  • В. А. Бондаренко
Keywords: ovarian tissue, 1, 2-propanediol, volume of oocytes, morphological structure


For the optimization of cryopreservation of ovarian tissue with the purpose of its using in clinical practice there have been carried out comparative analysis of the dynamics of volume and morphological transformation of the tissue at stepwise adding (1,5–3 M) of 1,2-propanediol (PROH) in various media. It has been shown that preservation of ovarian tissue structure and volume changes in the conditions of action of hyperosmolar solution of penetrating cryoprotective agent (CPA) are defined by the composition of the saturation solution and exposure time. At 10 minutes of tissues incubation in all studied cases there were observed no morphological changes in the volume and structure of tissue on the stage of saturation in PROH solutions. Increasing exposure time to 30 minutes led to cell compression to 40% in saturation medium containing 230 mM NaCl in 3 M PROH. The presence of fetal bovine serum (FBS) in similar conditions caused cells swelling both in 1.5 M concentration of PROH and 3 M. Oocytes volume remained within physiological values when using initial isotonic medium with increasing PROH concentration. It has been experimentally proved that the maximum morphological preservation of ovarian tissue after CPA saturation/removal is achieved at introduction of 10% FBS in hyperosmolar solutions of CPA.


Download data is not yet available.


Уикли Б. Электронная микроскопия для начинающих – М.: Мир, 1975. – 324с. /Uikli B. Elektronnaya mikroskopiya dlya nachinayushchikh– M.: Mir, 1975. – 324 s./

Amorim C.A., Rondina D., Rodrigues A. P.R. et al. Cryopreservation of isolated ovine primordial follicles with propylene glycol and glycerol // Fertility and Sterility. – 2004. – Vol.81, no 1. – P. 735–740.

Andersen C.Y., Rosendahl M., Byskov A.G. et al. Two successful pregnancies following autotransplantation of frozen/thawed ovarian tissue // Hum. Reprod. – 2008. – Vol.23, no 10. – P. 2266–2272.

Demeestere I., Simon P., Buxant F. et al. Ovarian function and spontaneous pregnancy after combined heterotopic and orthotopic cryopreserved ovarian tissue transplantation in a patient previously treated with bone marrow transplantation: case report // Hum. Reprod. – 2006a. – Vol.21. – P. 2010–2014.

Demeestere I., Simon P., Emiliani S. et al. Options to preserve fertility before oncological treatment: cryopreservation of ovarian tissue and its clinical application // Acta Clin. Belg. – 2006b. – Vol.61, no 5. – P. 259–263.

Demeestere I., Simon P., Emiliani S. et al. Orthotopic and heterotopic ovarian tissue transplantation // Hum. Reprod. Update. – 2009. – Vol.15, no 6. – P. 649–665.

Demirci B., Lornage J., Salle B. et al. Follicular viability and morphology of sheep ovaries after exposure to cryoprotectant and cryopreservation with different freezing protocols // Fertility and Sterility. – 2001. – Vol.75. – P. 754–762.

Donnez J., Dolmans M.M., Demylle D. et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue // Lancet. – 2004. – Vol.364. – P. 1405–1410.

Ghinea N., Fixman A., Alexandru D. et al. Identification of albumin-binding proteins in capillary endothelial cells // J. Cell Biol. – 1988. – Vol. 107, no 1. – P. 231–239.

Ghinea N., Eskenasy M., Simionescu M., Simionescu N. Endothelial albumin binding proteins are membrane-associated components exposed on the cell surface // J. Cell Biol. – 1989. – Vol.264. – P. 4755–4758.

Gougeon A. Dynamics of follicular growth in the human: a model from preliminary results // Hum. Reprod. – 1986. – No 1. – Р. 81–87.

Hreinsson J., Zhang P., Swahn M.L. et al. Cryopreservation of follicles in human ovarian cortical tissue. Comparison of serum and human serum albumin in the cryoprotectant solutions // Hum. Reprod. – 2003. – Vol.18. – P. 2420–2428.

Le Gal F., Gasqui P., Renard J.P. Differential osmotic behavior of mammalian oocytes before and after maturation: a quantitative analysis using goat oocytes as a model // Cryobiology. – 1994. – Vol.31. – P. 154–170.

Mazur P., Schneider U. Osmotic responses of preimplantation mouse and bovine embryos and their cryobiological implications // Cell Biophys. – 1986. – Vol.8. – P. 259–285.

Siflinger-Birnboim A., Malik A.B. Neutrophil adhesion to endothelial cells impairs the effects of catalase and glutathione in preventing endothelial injury // J. Cell Physiol. – 1993. – Vol.155, no 2. – P. 234–239.

Neto V., Buff S., Lornage J., Bottollier B. Effects of different freezing parameters on the morphology and viability of preantral follicles after cryopreservation of doe rabbit ovarian tissue // Fertil. Steril. – 2008. – Vol.89, no 5. – P. 1348–1356.

Newton H., Aubard Y., Rutherford A. et al. Low temperature storage and grafting of human ovarian tissue // Hum. Reprod. – 1996. – Vol.11. – P. 1487–1491.

Newton H., Pegg D.E., Barrass R., Gosden R.G. Osmotically inactive volume, hydraulic conductivity and permeability to dimethyl sulphoxide of human mature oocytes // Journal of Reproduction and Fertility. – 1999. – Vol.117. – P. 27–23.

Paynter S.J., Cooper A., Fuller B.J., Shaw R.W. Cryopreservation of bovine ovarian tissue: structural normality of follicles after thawing and culture in Vitro // Cryobiology. –1999. – Vol.38. – P. 301–309.

Santos R.R., Hurk R.v.d., Rodrigues A.P.R. et al. Effect of cryopreservation on viability, activation and growth of in situ and isolated ovine early-stage follicles // Animal Reproduction Science. – 2007. – Vol.99. – P. 53–64.

Schnitzer J.E., Bravo J. High affinity binding, endocytosis and degradation of conformationally-modified albumins: Potential role of gp30 and gp18 as novel scavenger receptors // J. Biol. Chem. – 1993. – Vol.268. – P. 7562–7570.

Schubert B., Canis M., Darcha C. et al. Human ovarian tissue from cortex surrounding benign cysts: a model to study ovarian tissue cryopreservation // Human Reproduction. – 2005. – Vol.20, no 7. – P. 1786–1792.

Shepard J.M., Goderie S.K., Brzyski N. et al. Effects of alterations in endothelial cell volume on transendothelial albumin permeabiIity // Journal of Cellular Physiology. – 1987. – Vol.133. – P. 389–394.

Songsasen N., Ratterree M.S., VandeVoort C.A. et al. Permeability characteristics and osmotic sensitivity of rhesus monkey (Macaca mulatta) oocytes // Human Reproduction. – 2002. – Vol.17, no 7. – P. 1875–1884.

Wang L., Liu J., Zhou G.-B. et al. Quantitative Investigations on the effects of exposure durations to the combined cryoprotective agents on mouse oocyte vitrification procedures // Biology of Reproduction. – 2011. – Vol.85. – P. 884–894.

Woods E.J., Benson J.D., Agca Y., Critser J.K. Fundamental cryobiology of reproductive cells and tissues // Cryobiology. – 2004. – Vol.48. – P. 146–156.
How to Cite
Трутаєва, І. А., Кірошка, В. В., Гавас, А. А., & Бондаренко, В. А. (1). Importance of fetal bovine serum introduction in hyperosmolar solutions of 1,2-propanediol for preserving morphological integrity of ovarian tissue. The Journal of V.N.Karazin Kharkiv National University. Series «Biology», 27, 91-98. Retrieved from