Evaluating the long-term effect of space radiation on the reproductive normality of mammalian sperm preserved on the International Space Station

Wakayama, Sayaka; Ito, Daiyu; Kamada, Yuko; Shimazu, Toru; Suzuki, Tomoo; Nagamatsu, Aiko; Araki, Ryoko; Ishikawa, Takahiro; Kamimura, Satoshi; Hirose, Naoki; Kazama, Kousuke; Li, Yang; Inoue, Rei; Kikuchi, Yasuyuki; Hayashi, Erika; Emura, Rina; Watanabe, Rie; Nagatomo, Hiroaki; Suzuki, Hiromi; Yamamori, Tohru; Tada, Motohide; Osada, Ikuko; Umehara, Masumi; Sano, Hiromi; Kasahara, Haruo; Higashibata, Akira; Yano, Sachiko; Abe, Masumi; Kishigami, Satoshi; Kohda, Takashi; Ooga, Masatoshi; Wakayama, Teruhiko

doi:10.1126/sciadv.abg5554

Cited by 28 publications

(18 citation statements)

References 51 publications

(80 reference statements)

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“…Determining whether mammalian species can reproduce normally in space or on other planets, where the environment markedly differs from that on Earth, including high levels of space radiation [ 1 , 2 ] and microgravity [ 3 ], is of particular interest. The International Space Station (ISS) is currently the only option for studying the actual effects of the space environment on mammalian reproduction; however, long-term breeding of mice is impossible [ 4 ].…”

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Mouse <i>in vivo</i>-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

et al. 2022

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Mammalian embryos are most commonly cryopreserved in liquid nitrogen; however, liquid nitrogen is not available in special environments, such as the International Space Station (ISS), and vitrified embryos must be stored at −80 °C. Recently, the high osmolarity vitrification (HOV) method was developed to cryopreserve mouse 2-cell stage embryos at −80 °C; however, the appropriate embryo is currently unknown. In this study, we compared the vitrification resistance of in vivo-derived, in vitro fertilization (IVF)-derived, and intracytoplasmic sperm injection (ICSI)-derived mouse 2-cell embryos against cryopreservation at −80 °C. The ICSI embryos had lower survival rates after warming and significantly lower developmental rates than the in vivo and IVF embryos. Further, IVF embryos had a lower survival rate after warming, but a similar rate to the in vivo embryos to full-term development. This result was confirmed by simultaneous vitrification of in vivo and IVF embryos in the same cryotube using identifiable green fluorescent protein-expressing embryos. We also evaluated the collection timing of the in vivo embryos from the oviduct and found that late 2-cell embryos had higher survival and developmental rates to full-term than early 2-cell embryos. Some early 2-cell embryos remained in the S-phase, whereas most late 2-cell embryos were in the G2-phase, which may have affected the tolerance to embryo vitrification. In conclusion, when embryos must be cryopreserved under restricted conditions, such as the ISS, in vivo fertilized embryos collected at the late 2-cell stage without long culture should be employed.

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Mouse <i>in vivo</i>-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

et al. 2022

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“…As a solution to this problem, we previously developed a freeze-dried storage method of mouse spermatozoa [ 16 ]. Using this method, freeze-dried spermatozoa can be readily transported at room temperature even between the Earth and International Space Station [ 17 , 18 ]. However, a method for the freeze-drying of embryos has not yet been developed.…”

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confidence: 99%

Optimised CO2-containing medium for in vitro culture and transportation of mouse preimplantation embryos without CO2 incubator

et al. 2021

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Conventional in vitro culture and manipulation of mouse embryos require a CO2 incubator, which not only increases the cost of performing experiments but also hampers the transport of embryos to the other laboratories. In this study, we established and tested a new CO2 incubator-free embryo culture system and transported embryos using this system. Using an Anaero pouch, which is a CO2 gas-generating agent, to increase the CO2 partial pressure of CZB medium to 4%–5%, 2-cell embryos were cultured to the blastocyst stage in a sealed tube without a CO2 incubator at 37°C. Further, the developmental rate to blastocyst and full-term development after embryo transfer were comparable with those of usual culture method using a CO2 incubator (blastocyst rate: 97% versus 95%, respectively; offspring rate: 30% versus 35%, respectively). Furthermore, using a thermal bottle, embryos were reliably cultured using this system for up to 2 days at room temperature, and live offspring were obtained from embryos transported in this simple and very low-cost manner without reducing the offspring rate (thermal bottle: 26.2% versus CO2 incubator: 34.3%). This study demonstrates that CO2 incubators are not essential for embryo culture and transportation and that this system provides a useful, low-cost alternative for mouse embryo culture and manipulation.

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“…We need mammalian sperm preservation technology in many fields, for example, maintenance of a huge mouse strains that increase every day. Freeze-drying technique is one of the methods to preserve the mammalian sperm ( Wakayama and Yanagimachi, 1998 ) and do not use liquid nitrogen (LN 2 ) or ultra-deep freezer during preservation ( Kamada et al., 2018 ; Wakayama et al., 2021 ). However, the conventional method using ampoules was not suitable for storing or transporting because the ampoules were made with glass and they were breakable, expensive, and bulky.…”

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confidence: 99%

Protocol to preserve mouse freeze-dried spermatozoa in the thin plastic sheets

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Wakayama

2021

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Evaluating the long-term effect of space radiation on the reproductive normality of mammalian sperm preserved on the International Space Station

Cited by 28 publications

References 51 publications

Mouse <i>in vivo</i>-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

Mouse <i>in vivo</i>-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

Optimised CO2-containing medium for in vitro culture and transportation of mouse preimplantation embryos without CO2 incubator

Protocol to preserve mouse freeze-dried spermatozoa in the thin plastic sheets

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