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Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa

OBJECTIVES: To challenge a vapour fast freezing (VFF) cryopreservation procedure (conventional VFF) with several vitrification protocols and VFF conducted with small semen volumes (10 μl, microVFF), in order to implement a procedure for sperm banking in subjects with small sperm number. MATERIALS AN...

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Autores principales: Arciero, Valentina, Ammar, Oumaima, Maggi, Mario, Vignozzi, Linda, Muratori, Monica, Dabizzi, Sara
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9544568/
https://www.ncbi.nlm.nih.gov/pubmed/35712876
http://dx.doi.org/10.1111/andr.13208
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author Arciero, Valentina
Ammar, Oumaima
Maggi, Mario
Vignozzi, Linda
Muratori, Monica
Dabizzi, Sara
author_facet Arciero, Valentina
Ammar, Oumaima
Maggi, Mario
Vignozzi, Linda
Muratori, Monica
Dabizzi, Sara
author_sort Arciero, Valentina
collection PubMed
description OBJECTIVES: To challenge a vapour fast freezing (VFF) cryopreservation procedure (conventional VFF) with several vitrification protocols and VFF conducted with small semen volumes (10 μl, microVFF), in order to implement a procedure for sperm banking in subjects with small sperm number. MATERIALS AND METHODS: Conventional VFF was conducted with test yolk buffer (TYB) as freezing medium and 500 μl straws as carriers. MicroVFF was conducted with TYB and using tips or cell sleepers as carriers. Vitrification was performed with TYB or SpermFreeze as freezing medium and with microspheres and tips as carriers. The effect of different procedures on progressive and total motility, viability, oxidative stress and DNA fragmentation of spermatozoa (sDF) was determined. Fresh and thawed samples, the latter after adequate washing/centrifuging, were evaluated. In some experiments, motility and viability recovery was determined in thawed samples, omitting the washing/centrifuging step. RESULTS: All the cryopreservation procedures blunted sperm motility and viability and induced increase of oxidative stress and sDF. However, VFF better preserved sperm motility and viability and less induced oxidative stress and sDF than vitrification, independently from the freezing medium and the carriers used in the latter. MicroVFF with cell sleepers resulted in a percentage increase of 57.58 ± 63.63%, 48.82 ± 74.96% and 24.55 ± 39.20% of, respectively, progressive and total motility and viability compared to the conventional VFF. Further, when tips were used, microVFF resulted in a percentage decrease of 15.77 ± 20.77% of sDF with respect to conventional VFF. Finally, omission of washing/centrifuging in post thawed samples, resulted in a much lower negative effect on motility and viability. DISCUSSION AND CONCLUSION: VFF, and in particular microVFF, better prevents sperm cryodamage than vitrification. Washing/centrifuging step after sample thawing seems to be responsible for a relevant fraction of damage to sperm motility and viability. Overall, our results are promising for developing a novel strategy of sperm banking in subjects with small sperm number, where low semen volumes are mandatory.
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spelling pubmed-95445682022-10-14 Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa Arciero, Valentina Ammar, Oumaima Maggi, Mario Vignozzi, Linda Muratori, Monica Dabizzi, Sara Andrology Original Articles OBJECTIVES: To challenge a vapour fast freezing (VFF) cryopreservation procedure (conventional VFF) with several vitrification protocols and VFF conducted with small semen volumes (10 μl, microVFF), in order to implement a procedure for sperm banking in subjects with small sperm number. MATERIALS AND METHODS: Conventional VFF was conducted with test yolk buffer (TYB) as freezing medium and 500 μl straws as carriers. MicroVFF was conducted with TYB and using tips or cell sleepers as carriers. Vitrification was performed with TYB or SpermFreeze as freezing medium and with microspheres and tips as carriers. The effect of different procedures on progressive and total motility, viability, oxidative stress and DNA fragmentation of spermatozoa (sDF) was determined. Fresh and thawed samples, the latter after adequate washing/centrifuging, were evaluated. In some experiments, motility and viability recovery was determined in thawed samples, omitting the washing/centrifuging step. RESULTS: All the cryopreservation procedures blunted sperm motility and viability and induced increase of oxidative stress and sDF. However, VFF better preserved sperm motility and viability and less induced oxidative stress and sDF than vitrification, independently from the freezing medium and the carriers used in the latter. MicroVFF with cell sleepers resulted in a percentage increase of 57.58 ± 63.63%, 48.82 ± 74.96% and 24.55 ± 39.20% of, respectively, progressive and total motility and viability compared to the conventional VFF. Further, when tips were used, microVFF resulted in a percentage decrease of 15.77 ± 20.77% of sDF with respect to conventional VFF. Finally, omission of washing/centrifuging in post thawed samples, resulted in a much lower negative effect on motility and viability. DISCUSSION AND CONCLUSION: VFF, and in particular microVFF, better prevents sperm cryodamage than vitrification. Washing/centrifuging step after sample thawing seems to be responsible for a relevant fraction of damage to sperm motility and viability. Overall, our results are promising for developing a novel strategy of sperm banking in subjects with small sperm number, where low semen volumes are mandatory. John Wiley and Sons Inc. 2022-06-27 2022-09 /pmc/articles/PMC9544568/ /pubmed/35712876 http://dx.doi.org/10.1111/andr.13208 Text en © 2022 The Authors. Andrology published by Wiley Periodicals LLC on behalf of American Society of Andrology and European Academy of Andrology. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Articles
Arciero, Valentina
Ammar, Oumaima
Maggi, Mario
Vignozzi, Linda
Muratori, Monica
Dabizzi, Sara
Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa
title Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa
title_full Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa
title_fullStr Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa
title_full_unstemmed Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa
title_short Vapour fast freezing with low semen volumes can highly improve motility and viability or DNA quality of cryopreserved human spermatozoa
title_sort vapour fast freezing with low semen volumes can highly improve motility and viability or dna quality of cryopreserved human spermatozoa
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9544568/
https://www.ncbi.nlm.nih.gov/pubmed/35712876
http://dx.doi.org/10.1111/andr.13208
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