Podufaly V. Cytomorphological and functional characteristics of cryopreserved spermatozoa with disorders of spermatogenesis.

Research objects were the processes of functional activity, morphological integrity, lipid peroxidation, DNA fragmentation, apoptosis and necrosis of native and cryopreserved human sperm at oligoasthenoteratozoospermia and azoospermia. The research aim was to study morphological characteristics of cryopreserved sperm obtained from the patients with azoospermia and oligoasthenoteratospermia, the influence of these characteristics on early embryogenesis and pregnancy rate in infertility treatment by the methods of assisted reproductive techniques. Research methods were cultural, cytological, morphometric, light microscopy, cryopreservation (programmable freezing), statistical ones. The results: In research we have found that during cryopreservation at oligoasthenoteratozoospermia the accumulation of spermatozoa with head structure pathology occurs.There was demonstrated an increase apoptosis and necrosis level of human spermatozoa from pathospermic ejaculates after cryopreservation. Elevated level of lipid peroxidation products - MDA indicates the presence of oxidative stress in spermatozoa in men with oligoasthenoteratozoospermia, which level is increased after the procedure of cryopreservation and reaches maximum values after accumulation of spermatozoa for future oocyte fertilization in vitro. There was established that level of DNA fragmentation, apoptosis and necrosis in ejaculatory spermatozoa was significantly higher than in testiculatoty ones. These findings can be used to formulate the strategies to overcome male factor in infertile couples. In this regard, we propose a new approach to select the treatment strategy for couples with disorders of spermatogenesis: the choice of single morphologically normal spermatozoa and their introduction into oocyte cytoplasm. New data: For the first time there were systematically investigated the morphological characteristics of cryopreserved human spermatozoa isolated from ejaculates of men who have had severe disordered spermatogenesis, namely in oligoasthenoteratospermia and testicular biopsy in the patients with azoospermia. For the first time it has been revealed that in the process of cryopreservation there is an accumulation of spermatozoa with head pathology, requiring an additional selection of cryopreserved sperm according to morphological criteria. Increased lipid peroxidation, apoptosis and necrosis, DNA fragmentation in human spermatozoa after cryopreservation point to the need of using the technique of intracytoplasmic injection of morphologically selected spermatozoon to prevent possible abnormal fertilization.