Primerjava učinkovitosti tehnologij ultrahitrega in dvostopenjskega zamrzovanja semena moških v tekočem dušiku.

Kovač, Katarina (author), Kovačič, Borut (mentor)

V raziskavi smo ugotavljali spremembo gibljivosti semenčic pred in po dvostopenjskim in ultrahitrim zamrzovanjem ter odtajanjem semenskega vzorca. Prospektivna primerjalna študija je bila izvedena na 27 svežih vzorcih semena, od tega na 15 vzorcih semena moških z normozoospermijo in 12 z astenozoospermijo. Posamezen semenski vzorec je bil pred zamrzovanjem razdeljen na 4 dele. Dva dela smo razredčili s klasičnim krioprotektantom, ki je vseboval glicerol (Sperm Freeze Medium), druga dva dela pa s krioprotektantom in dodanim jajčnim rumenjakom (TEST-yolk buffer). Iz vsake od obeh skupin smo polovico tako pripravljenega semena zamrznili po ultrahitri metodi v mikrokapljicah, ostalo polovico pa po dvostopenjski metodi v slamicah. Gibljivost semenčic smo ocenili z računalniškim programom. Ugotavljali smo, ali bo po odmrzovanju gibljivost semenčic statistično podobna v primeru ultrahitrega in dvostopenjskega zamrzovanja in kako na to vpliva prisotnost oziroma odsotnost jajčnega rumenjaka v mediju za zamrzovanje. Rezultati so pokazali enako stopnjo gibljivosti in progresivne gibljivosti semenčic ter enako hitrost vijugaste poti v primeru obeh metod zamrzovanja, ne glede na to, kateri krioprezervacijski medij smo uporabili. Nova metoda zamrzovanja semena v mikrokapljicah je uporabna za zamrzovanje majhnih volumnov semenskega vzorca, tudi ob poslabšani gibljivosti semenčic

• Language: Slovenian
• Keywords: reproduktivna medicina, gibljivost semenčic, ultrahitro zamrzovanje, dvostopenjsko zamrzovanje
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [K. Kovač]
• Year: 2018
• PID: 20.500.12556/RUL-101589
• UDC: 606:611.63.013.1:57.086.13(043.2)
• COBISS.SI-ID: 8993401
• Publication date in RUL: 20.06.2018

Secondary language

• Language: English
• Title: Comparison of ultrarapid and two-step freezing of human semen in liquid nitrogen

Abstract

In a research we were analysing the motility of human spermatozoa before and after two-step and ultrarapid cryopreservation of semen sample. A prospective comparative study was performed on 27 fresh semen samples. From this group, 15 men were diagnosed with normozoospermia and 12 with asthenozoospermia. Each semen sample was divided on 4 aliquotes. First two aliquotes were diluted with glycerol medium (Sperm Freeze Medium). Another two aliquotes were diluted with TEST-yolk buffer, which contains glycerol and egg yolk. In every group, one aliquot was frozen with two-step method and another one with ultrarapid method. After thawing, the motility of spermatozoa was evaluated with computer analysis. The main question was if ultrarapid freezing gives statistically similar results in postthaw motility comparing to two-step cryopreservation method. Results have shown the same level of postthaw motility and progressive motility of spermatozoa. There was also the same level of curvilinear velocity after both methods of cryopreservation and after the use of both cryoprotectants. Ultrarapid cryopreservation of microdrops could be appropriate method for freezing small volumes of semen sample, even if the motility of spermatozoa is low.

• Keywords: reproductive medicine, sperm motility, ultrarapid freezing, two-step freezing