Poor sperm quality is a common cause of male infertility. Its fertilizing ability is determined by morphological and ultrastructural properties and membrane integrity, while embryo development, quality and pregnancy outcome are greatly influenced by chromatin maturity, structure and DNA damage. In this master thesis, we compared two sperm preparation techniques using double-layer gradient centrifugation vs. microfluidics. Sorting with microfluidics simulates obstacles similar to the cervix and uterus, mimicking natural sperm selection. When examining sperm quality, we evaluated hyaluronan binding, morphology, nucleoprotein maturity and DNA fragmentation. The proportions of hyaluronan-binding sperm and morphologically normal sperm were comparable in both methods. The proportions of mature nucleoproteins and DNA fragmentation indicate that the microfluidic sperm preparation method is more successful in selecting sperm with higher genetic integrity. After fertilization with sperm prepared with DGC and MF, we checked the proportions, we review the percentage of fertilized cells, the quality of the embryos and the pregnancy rate depending on the preparation method. A review of the outcome of in vitro fertilization procedures of treated couples shows that sperm preparation using microfluidics and centrifugation is comparable. The outcome is also comparable in the group where the woman age is above 38 and where the spermatozoa concentration of the man is less than 5×10^6/mL. The method is not suitable for preparing sperm with a concentration of spermatozoa less than or equal to 1×10^6/mL. There is a possible correlation between the microfluidic method and the age of women over 40, as sperm selection has a lower percentage of DNA fragmentation index and a higher pregnancy rate, as well as a lower ability of the oocyte to repair DNA.
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