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

Abstract

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.