Hapon H. Structural and functional state of single human sperm after cryopreservation

The object of the study is changes in the morphofunctional and ultrastructural characteristics of sperm of men with OAT after cryopreservation with high molecular weight PVP solution. The purpose of the study - to determine the ultrastructural and morphofunctional characteristics of single human sperm in oligoastenoteratozoospermia after cryopreservation. Research methods: light microscopy to determine the morphological characteristics of human sperm; electron microscopy to assess the structure of their ultrastructural elements. To study the functional characteristics of sperm before and after cryopreservation, their motility was assessed using computer analysis CASA; used SCD test to determine the level of DNA fragmentation; flow cytofluorimetry using staining with a potential-dependent probe JC-1 to determine the number of sperm with high mitochondrial potential; cryobiological and statistical. In the work for the first time the method of cryopreservation of spermatozoa of men with defects of spermatogenesis with 10% of PVP (m. 360000) as a cryoprotectant was developed. It is shown that the indicators of motility, viability and fertilizing ability of sperm cryopreserved with PVP do not differ significantly from those for native cells. For the first time the morphological and ultrastructural characteristics of spermatozoa after cryopreservation with PVP were analyzed. The frequency of DNA fragmentation, the number of cells cryopreserved with PVP with high mitochondrial potentialwere determined. The indicators of oocyte fertilization by spermatozoa cryopreserved with PVP were studied and morphokinetic analysis of embryo development in vitro was performed.