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Petrushko M, Yurchuk T, Todorov P, et al. New method for cryoprotectant-free freezing of human oligoasthenoteratozoospremic spermatozoa with high-molecular polymer. Cryobiology. 2021;doi: 10.1016/j.cryobiol.2021.09.013.
Petrushko, M., Yurchuk, T., Todorov, P., Hristova, E., Piniaiev, V., Isachenko, E., Rahimi, G., Mallmann, P., & Isachenko, V. (2021). New method for cryoprotectant-free freezing of human oligoasthenoteratozoospremic spermatozoa with high-molecular polymer. Cryobiology, . https://doi.org/10.1016/j.cryobiol.2021.09.013
Petrushko, Maryna, et al. "New method for cryoprotectant-free freezing of human oligoasthenoteratozoospremic spermatozoa with high-molecular polymer." Cryobiology vol. (2021). doi: https://doi.org/10.1016/j.cryobiol.2021.09.013
Petrushko M, Yurchuk T, Todorov P, Hristova E, Piniaiev V, Isachenko E, Rahimi G, Mallmann P, Isachenko V. New method for cryoprotectant-free freezing of human oligoasthenoteratozoospremic spermatozoa with high-molecular polymer. Cryobiology. 2021 Oct 02; doi: 10.1016/j.cryobiol.2021.09.013. Epub 2021 Oct 02. PMID: 34606824.
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Cryobiology. 2021 Oct 02; doi: 10.1016/j.cryobiol.2021.09.013. Epub 2021 Oct 02.

New method for cryoprotectant-free freezing of human oligoasthenoteratozoospremic spermatozoa with high-molecular polymer.

Cryobiology

Maryna Petrushko, Taisiia Yurchuk, Plamen Todorov, Elena Hristova, Volodymyr Piniaiev, Evgenia Isachenko, Gohar Rahimi, Peter Mallmann, Vladimir Isachenko

Affiliations

  1. Institute for Problems of Cryobiology and Cryomedicine, 61016, Kharkiv, Ukraine; ART-clinic of Reproductive Medicine, 61000, Kharkiv, Ukraine.
  2. Institute for Problems of Cryobiology and Cryomedicine, 61016, Kharkiv, Ukraine.
  3. Institute of biology and Immunology of Reproduction, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria.
  4. ART-clinic of Reproductive Medicine, 61000, Kharkiv, Ukraine.
  5. Research Group for Reproductive Medicine, Medical Faculty, Department of Obstetrics and Gynaecology, Cologne University, 50931, Cologne, Germany.
  6. Research Group for Reproductive Medicine, Medical Faculty, Department of Obstetrics and Gynaecology, Cologne University, 50931, Cologne, Germany. Electronic address: [email protected].

PMID: 34606824 DOI: 10.1016/j.cryobiol.2021.09.013

Abstract

Data about cryoprotectant-free cryopreservation of human ICSI spermatozoa are limited. The aim of this investigation was to compare two technologies for cryopreservation of spermatozoa from men with oligoasthenoteratozoospermia: standard conventional freezing with 5% glycerol (freezing in glycerol) and cryoprotectant-free freezing with 5% high-molecular-weight (360 kDa) polyvinylpyrrolidone (PVP) (PVP-freezing). Capillaries with spermatozoa were cooled in vapor and then plunginged into liquid nitrogen. Head-, midpiece- and tail-abnormality of spermatozoa, mitochondrial membrane potential (MMP) and DNA fragmentation rates after cryopreservation were evaluated. After warming of spermatozoa, fertilization of oocytes (ICSI) was performed. It was detected the lower rate of morphological abnormalities of PVP-frozen spermatozoa in comparison with cells frozen with glycerol (34.6 ± 4.1% vs. 20.7 ± 4.7%, respectively) (P < 0.05). Quality of cells with high MMP after warming in spermatozoa frozen with glycerol was lower than in PVP-frozen spermatozoa (34.7 ± 4.2 vs. 54.5 ± 4.2%, respectively) (P < 0.05). It was established that the DNA fragmentation rate in PVP-frozen spermatozoa was significantly lower in comparison with spermatozoa frozen with glycerol (23.1 ± 2.5% vs. 38.8 ± 3.0%, respectively) (P < 0.05). After fertilization (ICSI) of oocytes, it was established that cleavage and blastulation rates were higher in oocytes after fertilization with PVP-frozen spermatozoa than with spermatozoa frozen with glycerol. Fertilization-, development to 8-blastomeres-, and blastocyst-rates were for PVP-frozen and spermatozoa frozen with glycerol, respectively: 94.4 ± 7.8 vs. 82.2 ± 6.2% (P > 0.1 with tendency to increasing), 90.0 ± 4.6 vs. 69.5 ± 5.1% (P < 0.05), and 45.4 ± 4.1% vs. 30.9 ± 3.3% (P < 0.05). It was concluded that permeable cryoprotectant-free freezing with 5% high-molecular-weight (360 kDa) polyvinylpyrrolidone can be applied successfully for cryopreservation of human oligoasthenoteratozoospremic spermatozoa.

Copyright © 2021 Elsevier Inc. All rights reserved.

Keywords: Cryopreservation; Embryo development; Freezing; Glycerol; Oligoasthenoteratozoospermia; Polyvinylpyrrolidone

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