Abstract
ObjectiveCryopreservation of human spermatozoa is fundamental in assisted reproductive technology. At present, slow freezing techniques are widely used for sperm cryopreservation. Recently, sperm vitrification has been proposed as an alternative to slow freezing. This study aimed to compare the efficiency of slow versus ultra-rapid freezing after thawing and to determine the level of DNA fragmentation in post-thaw normal human semen samples processed through each of the cryopreservation techniques.MethodsUltra-rapid freezing is a method that only differs from conventional ultra-rapid freezing in the use of sucrose as a cryoprotectant. In experiment 1, 24 semen samples were used to compare sperm recovery rates after slow and ultra-rapid sperm freezing. In experiment 2, 18 semen samples were used to compare post-thaw sperm DNA fragmentation levels after each of the cryopreservation techniques.ResultsIn experiment 1, no significant differences were observed in sperm concentration recovery rates, although slow freezing showed a lower progressive motility rate than ultra-rapid freezing (16.6±7.4% vs. 34.7±10.2%), and higher non-progressive and immotile sperm counts (9.0±4.0% vs. 7.6±2.8%; and 74.4±10.1% vs. 57.8±10.3%, respectively). In experiment 2, sperm DNA fragmentation after thawing was significantly higher in slow freezing than in fresh post gradient processing and ultra-rapid freezing samples (47.3±13.4% vs. 9.1±3.7% vs. 14.6±4.6%, respectively).ConclusionSperm ultra-rapid freezing may be an alternative to slow freezing with better recovery results and less apparent DNA damage.
Highlights
IntroductionCryopreservation of human spermatozoa is a fundamental resource in assisted reproductive technology (ART) that allows the optimization of infertility treatment and male fertility preservation therapy prior to chemotherapy, radiotherapy or testicular radical surgery (Sanger et al, 1992)
In experiment 1, no significant differences were observed in sperm concentration recovery rates, slow freezing showed a lower progressive motility rate than ultra-rapid freezing (16.6±7.4% vs. 34.7±10.2%), and higher non-progressive and immotile sperm counts (9.0±4.0% vs. 7.6±2.8%; and 74.4±10.1% vs. 57.8±10.3%, respectively)
Cryopreservation of human spermatozoa is a fundamental resource in assisted reproductive technology (ART) that allows the optimization of infertility treatment and male fertility preservation therapy prior to chemotherapy, radiotherapy or testicular radical surgery (Sanger et al, 1992)
Summary
Cryopreservation of human spermatozoa is a fundamental resource in assisted reproductive technology (ART) that allows the optimization of infertility treatment and male fertility preservation therapy prior to chemotherapy, radiotherapy or testicular radical surgery (Sanger et al, 1992). Slow freezing techniques have been widely used in sperm cryopreservation, allowing the storage of large sample volumes with acceptable results for sperm vitality and motility after thawing (Fuller et al, 2004; Donnez & Kim, 2011). Vitrification has proven its effectiveness from oocytes to embryos These have been possible due to an acceleration of the cryopreservation process by minimizing the volumes of the solution with the development of different vitrification devices and the optimization of the cryoprotectant combination. The method described by Isachenko et al (2003; (2004; 2008) and Isachenko et al (2004;2005) for sperm vitrification only differs from the technique developed for oocytes and embryos in the use of sucrose as the only cryoprotectant
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