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Cryopreservation of very low numbers of spermatozoa from male patients undergoing infertility treatment using agarose capsules

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Abstract

This study tried to cryopreserve low numbers of spermatozoa from men undergoing infertility treatments by inserting into agarose capsules. The capsules were transferred into a drop of cryoprotectant solution and injected 3–4 motile spermatozoa that were selected by the swim-up method by conventional intracytoplasmic sperm injection. These capsules were put on a Cryotop® and frozen in liquid nitrogen vapor, and then submerged into liquid nitrogen and subsequently thawed and recovered. The motile spermatozoa in the capsules were counted. Eventually, we cryopreserved 2142 motile spermatozoa in 702 agarose capsules from 26 male patients and 1356 (63%) spermatozoa maintained their motility after thawing. The spermatozoa motility rates after thawing (MRAT) ranged from 20.0% (5/25) to 95.1% (58/61) among patients. The median MRAT was 68.3% (interquartile range 46.1–75.7). The total number of motile spermatozoa collected by swim-up method strongly correlated with MRAT (r = 0.746). It was possible to cryopreserve spermatozoa from male patients undergoing infertility treatment using agarose capsules. However, there were wide differences in MRAT among patients. It seems the spermatozoa from semen where there were many motile spermatozoa may have higher freezing resistance. Further studies using this method in cryptozoospermic semen, testicular and epididymal spermatozoa are required.

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Acknowledgements

We would like to thank Louise Kellam, Deputy Lab Manager at CARE Fertility, Nottingham, UK, for assistance with English language editing. We are also grateful to Mr. Satoshi Sakahara at the Graduate School of Science and Engineering, Yamagata University, for assistance with scanning electron microscopy. Also, we thank Dr. Kimura at Gumma Paz College for statistical analysis advice.

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Author notes

  1. Shota Hatakeyama

    Present address: Yanaihara Women’s Clinic, 1-26-29 Ofuna, Kamakura, 247-0056, Japan

Authors and Affiliations

  1. Tokuoka Ladies Clinic, 1-3-1 Nakane, Meguro-Ku, Tokyo, 152-0031, Japan

    Shota Hatakeyama & Susumu Tokuoka

  2. Graduate School of Health Science, Gumma Paz College, 1-7-1 Tonyamachi, Takasaki, 370-0006, Japan

    Shota Hatakeyama & Yasuhisa Araki

  3. Graduate School of Science and Engineering, Yamagata University, 4-3-16 Jonan, Yonezawa, 992-8510, Japan

    Hiroyuki Abe

  4. The Institute for Advanced Reproductive Medical Technology, 909-21 Ishi, Fujimi-Cho, Maebashi, 371-0105, Japan

    Yasuyuki Araki & Yasuhisa Araki

  5. Nippon Reprogenetics, Maebashi, 909-21 Ishi, Fujimi-Cho, Maebashi, 371-0105, Japan

    Yasuhisa Araki

Authors
  1. Shota Hatakeyama
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  2. Susumu Tokuoka
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  4. Yasuyuki Araki
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Correspondence to Shota Hatakeyama.

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The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in this study involving human participants were in accordance with the ethical standards of the national committee, and were approved by the Ethics Committee at Gumma Paz College and Tokuoka Ladies Clinic.

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Informed consent was obtained from all individual participants included in the study.

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Hatakeyama, S., Tokuoka, S., Abe, H. et al. Cryopreservation of very low numbers of spermatozoa from male patients undergoing infertility treatment using agarose capsules. Human Cell 30, 201–208 (2017). https://doi.org/10.1007/s13577-017-0166-x

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  • DOI: https://doi.org/10.1007/s13577-017-0166-x

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