Birth of mice from vitrified/warmed 2-cell embryos transported at a cold temperature

Tanaka, Takeo; Kaneko, Takehito; Haruguchi, Yukie; Fukumoto, Kiyoko; Machida, Hiromi; Koga, Mika; Nakagawa, Yoshiko; Takeshita, Yumi; Matsuguma, Toyokazu; Tsuchiyama, Shuuji; Shimizu, Norihiko; Hasegawa, Takanori; Goto, Motohito; Miyachi, Hitoshi; Anzai, Masayuki; Nakatsukasa, Ena; Nomaru, Koji; Nakagata, Naomi

doi:10.1016/j.cryobiol.2008.12.011

Cited by 22 publications

(21 citation statements)

References 18 publications

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“…This demonstrates that successful transfer of morulae after 24 h of storage can be achieved in any of the media or temperature assayed without compromising in vivo viability. Previous studies have focused on the transport of 2-cell embryos, either thawed after cryopreservation, [30] or obtained from in vitro fertilization [5], achieving 44% in vivo development for embryos stored at 4°C for 48 hours. We obtained higher developmental levels under all the conditions tested, but in our study, embryo transfer was performed after 24 hours of storage.…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

Hourcade

Pérez-Crespo

Serrano

et al. 2012

Reprod Biol Endocrinol

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BackgroundInterchange of genetically modified (GM) mice between laboratories using embryos provides several advantages. Not only is transport stress avoided, but also the health status of the recipient colony is not compromised. Embryos do not need to be shipped in frozen stage, which requires expensive packaging in addition to a certain degree of expertise in order to freeze and thaw them correctly. The aim of this study was to examine different storage conditions and their effect on embryo viability in order to establish the feasibility of practical, non-frozen conditions for embryo shipment.MethodsMouse morulae developed in vivo (collected from donors 2.5d post coitum) or in vitro (zygotes cultured until morulae stage) were stored, combining two different media (KSOMeq or KSOM-H) and temperatures (4 degrees C, 15 degrees C and 37 degrees C) throughout 24 or 48 hours. After storage in vitro viability was assessed determining percentage of development to blastocyst and total cell number. In vivo viability was determined based on the number of implantations and living fetuses after embryo transfer of stored embryos. The storage effect at the molecular level was assessed by studying a gene pool involved in early development by quantitative RT-PCR.ResultsIn vivo-produced morulae stored for 24 hours did not show differences in development up to the blastocyst stage, regardless of the storage type. Even though a decrease in the total cell number in vivo was observed, embryo development after embryo transfer was not affected. All 24 hour storage conditions tested provided a similar number of implantations and fetuses at day 14 of pregnancy. Morulae obtained from in vitro embryo culture collected at the 1-cell stage showed a decreased ability to develop to blastocyst after 24 hours of storage at 15degrees C both in KSOMeq and KSOM-H. Concomitantly, a significant decrease of embryo implantation rates after transfer to recipients was also found. In order to further characterize the effect of non-frozen storage combining a molecular approach with the ordinary in vitro culture evaluation, embryos collected at the morula stage were submitted to the same storage conditions described throughout 48 hours. In vitro culture of those embryos showed a significant decrease in their developmental rate to blastocyst in both KSOMeq and KSOM-H at 15degrees C, which also affected the total number of cells. Gene transcription studies confirmed significant alterations in retrotransposons (Erv4 and Iap) after 48 h of storage at 15degrees C.ConclusionsOur results show that both KSOMeq and KSOM-H can be equally used, and that several temperature conditions allow good survival rates in vitro and in vivo. Some of these storage conditions can substitute freezing in order to maintain embryo viability for 24–48 hours, providing a reliable and less demanding technical alternative for embryo interchanges.

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Section: Discussionmentioning

confidence: 99%

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

Hourcade

Pérez-Crespo

Serrano

et al. 2012

Reprod Biol Endocrinol

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“…Of particular relevance to the mouse community is the ability to transport unfrozen embryos that have previously been cryopreserved. This allows laboratories that do not have access to LN 2 or are not familiar with handling frozen embryos to take advantage of the vast numbers of mouse strains held in archives around the world without resorting to live animal transportation (Takeo et al, 2009(Takeo et al, , 2010.…”

Section: Transporting Frozen/thawed Embryosmentioning

confidence: 99%

“…Unfrozen embryos may also be used to facilitate the exchange of mouse stocks between laboratories. Using this approach, embryos held at ß4°to 8°C have been exchanged between Japan, the U.K., and the U.S.A. (Takeo et al, 2009(Takeo et al, , 2010. Although this procedure offers some very significant cost savings, it is essential to use a reliable transport chain.…”

Section: Background Informationmentioning

confidence: 99%

Transporting Mouse Embryos and Germplasm as Frozen or Unfrozen Materials

et al. 2014

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The 21st century has seen a huge proliferation in the availability of genetically altered mice. The availability of these resources has been accompanied by ever greater opportunities for international collaborations between laboratories involving the exchange of mouse strains. This exchange can involve significant costs in terms of animal welfare and transportation expenses. In an attempt to mitigate some of these costs, the mouse community has developed a battery of techniques that can be used to avoid transporting live mice. Transporting frozen embryos and sperm at liquid nitrogen (LN2 ) temperatures using dry shippers has been common practice for some time. However, current advances in this field have refined transportation procedures and introduced new techniques for disseminating embryos and sperm: for example, shipping frozen sperm on dry ice, exchanging unfrozen epididymides from which sperm can be extracted, and transporting frozen/thawed embryos in isotonic media. This article discusses some of the current practices used by laboratories to transport mouse strains around the world without having to exchange live mice.

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“…In addition, cryoprotectant, preincubation medium, and fertilization medium for use in this method are available (http://www.kyudo.co.jp/Fertiup/FERTOPen.htm). CARD scientists also have demonstrated that frozen-thawed 2-cell embryos and 2-cell embryos produced by frozen-thawed sperm and transported from our center to many institutes at a refrigerated temperature have the ability to develop well into live young (Takeo et al 2009, Takeo et al 2010). The advantages of this method are that the recipients do not have to master thawing techniques, and special containers (e.g., dry shippers) are not required for embryo transport.…”

Section: Repository Profilesmentioning

confidence: 99%

Centralized mouse repositories

et al. 2012

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Because the mouse is used so widely for biomedical research and the number of mouse models being generated is increasing rapidly, centralized repositories are essential if the valuable mouse strains and models that have been developed are to be securely preserved and fully exploited. Ensuring the ongoing availability of these mouse strains preserves the investment made in creating and characterizing them and creates a global resource of enormous value. The establishment of centralized mouse repositories around the world for distributing and archiving these resources has provided critical access to and preservation of these strains. This article describes the common and specialized activities provided by major mouse repositories around the world.

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Birth of mice from vitrified/warmed 2-cell embryos transported at a cold temperature

Cited by 22 publications

References 18 publications

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

In vitro and in vivodevelopment of mice morulae after storage in non-frozen conditions

Transporting Mouse Embryos and Germplasm as Frozen or Unfrozen Materials

Centralized mouse repositories

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