Association between freezing agent and acrosome damage of human spermatozoa from subnormal and normal semen

This experimental study compares the effects of human sperm preservation medium (HSPM) with TEST-yolk buffer (TYB) as cryoprotectants of human spermatozoa with respect to the integrity of the acrosome after the freeze-thawing procedure. Fifty-six semen samples were included in this study; 18 were subnormal (G1) and 38 were normal (G2) based on World Health Organization criteria, except for morphology, which was evaluated according to strict criteria. Each semen sample was divided into two parts: the first part was prepared for cryopreservation by the addition of HSPM (1:1) and the second by addition of TYB (1:1). Freezing was performed in liquid nitrogen vapour. Smears were made before freezing and after the thawing process for evaluation of acrosome integrity using fluorescent-lectin labelling. The mean percentage of spermatozoa with intact acrosomes in the subnormal group was 77.0 +/- 7.2% before freezing and decreased significantly (P < 0.001) after thawing: to 63.7 +/- 8.2% with the use of HSPM and 66.8 +/- 8.7% with the use of TYB. The corresponding values in the normal semen samples were 83.4 +/- 9.2%, 76.0 +/- 8.8% and 77.9 +/- 9.2%, respectively. It is obvious that the decrease in the mean percentage of spermatozoa with intact acrosome was significantly higher when using HSPM in comparison with TYB, not only for G1 (-14.9 +/- 1.9% versus -11.8 +/- 1.4%) but also for G2 samples (-13.8 +/- 1.5% versus -11.9 +/- 1.3%). In conclusion, TYB should be recommended for freeze-thawing of human spermatozoa as the first-choice cryoprotectant, for normal as well as subnormal semen samples, in order to protect the sperm acrosome from the deleterious effects of the freeze-thawing procedure.