Introduction: This study compared the effects of three different media on human sperm parameters, and ultrastructure of spermatozoa using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) after vitirification. These media were artificial seminal fluid (ASF), seminal fluid (SF) and human tubal fluid (HTF)-sucrose. Materials and Methods: 30 normal ejaculates were processed with swim-up technique and sperm suspensions were divided in four aliquots: 1) fresh sample (control); 2) vitrification in HTF supplemented with 0.25 mol sucrose, as routine procedure; 3) vitrification with patients‟ SF; and 4) vitrification in ASF. After warming, sperm parameters of motility, viability and morphology were analyzed using WHO criteria. Also, sperm pellets were fixed in 2.5% gluthardehyde and processed for SEM and TEM observations. Results: Sperm parameters in all cryo-groups were reduced when compared with control samples (p<0.0001). Briefly, sperm grade A motility, viability and normal morphology were significantly higher in ASF than HTF group. After cryopreservation, deep invagination in cytoplasm, mechanically weak point sites, rough membrane surface and looped tails were observed in SEM evaluation. The looped tails were more severe in SF and HTF cryo-groups. In TEM evaluation, acrosome damage, plasma membrane loss, chromatin vacuolation, and disruption of mitochondria arrangement and structure were observed in all cryogroups. Degradation of cells was also observed, especially in HTF cryo-group. Conclusion: Vitrification of human spermatozoa with ASF can effectively preserve the quality of motility in comparison with routine procedure. This ASF medium formulated according to SF that is a natural medium for sperm preservation, lacking artificial components, such as sucrose. With this design, any osmotic shock can be deleted before and after freezing. Also, this study confirmed that SF in normal ejaculates can act as cryoprotectants.
Artificial seminal fluid preserves human sperm quality in vitrification program: An electron microscopy study / A., Agha Rahimi; M. A., Khalili; Nottola, Stefania Annarita; A. R., Moradi. - In: IRANIAN JOURNAL OF REPRODUCTIVE MEDICINE. - ISSN 1680-6433. - STAMPA. - 13, n° 4, suppl. 1:(2015), pp. 13-13. (Intervento presentato al convegno 6th Yazd International Congress and Student Award in Reproductive Medicine. tenutosi a Yazd, Iran nel April 17-19, 2015).
Artificial seminal fluid preserves human sperm quality in vitrification program: An electron microscopy study.
NOTTOLA, Stefania Annarita;
2015
Abstract
Introduction: This study compared the effects of three different media on human sperm parameters, and ultrastructure of spermatozoa using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) after vitirification. These media were artificial seminal fluid (ASF), seminal fluid (SF) and human tubal fluid (HTF)-sucrose. Materials and Methods: 30 normal ejaculates were processed with swim-up technique and sperm suspensions were divided in four aliquots: 1) fresh sample (control); 2) vitrification in HTF supplemented with 0.25 mol sucrose, as routine procedure; 3) vitrification with patients‟ SF; and 4) vitrification in ASF. After warming, sperm parameters of motility, viability and morphology were analyzed using WHO criteria. Also, sperm pellets were fixed in 2.5% gluthardehyde and processed for SEM and TEM observations. Results: Sperm parameters in all cryo-groups were reduced when compared with control samples (p<0.0001). Briefly, sperm grade A motility, viability and normal morphology were significantly higher in ASF than HTF group. After cryopreservation, deep invagination in cytoplasm, mechanically weak point sites, rough membrane surface and looped tails were observed in SEM evaluation. The looped tails were more severe in SF and HTF cryo-groups. In TEM evaluation, acrosome damage, plasma membrane loss, chromatin vacuolation, and disruption of mitochondria arrangement and structure were observed in all cryogroups. Degradation of cells was also observed, especially in HTF cryo-group. Conclusion: Vitrification of human spermatozoa with ASF can effectively preserve the quality of motility in comparison with routine procedure. This ASF medium formulated according to SF that is a natural medium for sperm preservation, lacking artificial components, such as sucrose. With this design, any osmotic shock can be deleted before and after freezing. Also, this study confirmed that SF in normal ejaculates can act as cryoprotectants.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
