High Osmolality Vitrification: A New Method for the Simple and Temperature-Permissive Cryopreservation of Mouse Embryos

Keiji Mochida, Ayumi Hasegawa, Ming Wen Li, Martin D. Fray, Seiji Kito, Jadine M. Vallelunga, Kevin C K Lloyd, Atsushi Yoshiki, Yuichi Obata, Atsuo Ogura

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Procedures for cryopreserving embryos vary considerably, each having its specific advantages and disadvantages in terms of technical feasibility, embryo survival yield, temperature permissibility and species- or strain-dependent applicability. Here we report a high osmolality vitrification (HOV) method that is advantageous in these respects. Cryopreservation by vitrification is generally very simple, but, unlike slow freezing, embryos should be kept at a supercooling temperature (below -130°C) to avoid cryodamage. We overcame this problem by using an HOV solution containing 42.5% (v/v) ethylene glycol, 17.3% (w/v) Ficoll and 1.0 M sucrose. This solution is more viscous than other cryopreservation solutions, but easy handling of embryos was assured by employing a less viscous equilibration solution before vitrification. Most (>80%) embryos cryopreserved in this solution survived at -80°C for at least 30 days. Normal mice were recovered even after intercontinental transportation in a conventional dry-ice package for 2-3 days, indicating that special containers such as dry shippers with liquid nitrogen vapor are unnecessary. The HOV solution could also be employed for long-term storage in liquid nitrogen, as with other conventional cryoprotectants. Finally, we confirmed that this new vitrification method could be applied successfully to embryos of all six strains of mice we have tested so far. Thus, our HOV method provides an efficient and reliable strategy for the routine cryopreservation of mouse embryos in animal facilities and biomedical laboratories, and for easy and cheap transportation.

Original languageEnglish (US)
Article numbere49316
JournalPLoS One
Volume8
Issue number1
DOIs
StatePublished - Jan 16 2013

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Vitrification
Cryopreservation
vitrification
osmolality
cryopreservation
Osmolar Concentration
embryo (animal)
Embryonic Structures
Temperature
mice
temperature
Liquid nitrogen
methodology
Nitrogen
Dry Ice
ficoll
Supercooling
Ficoll
supercooling
liquids

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Mochida, K., Hasegawa, A., Li, M. W., Fray, M. D., Kito, S., Vallelunga, J. M., ... Ogura, A. (2013). High Osmolality Vitrification: A New Method for the Simple and Temperature-Permissive Cryopreservation of Mouse Embryos. PLoS One, 8(1), [e49316]. https://doi.org/10.1371/journal.pone.0049316

High Osmolality Vitrification : A New Method for the Simple and Temperature-Permissive Cryopreservation of Mouse Embryos. / Mochida, Keiji; Hasegawa, Ayumi; Li, Ming Wen; Fray, Martin D.; Kito, Seiji; Vallelunga, Jadine M.; Lloyd, Kevin C K; Yoshiki, Atsushi; Obata, Yuichi; Ogura, Atsuo.

In: PLoS One, Vol. 8, No. 1, e49316, 16.01.2013.

Research output: Contribution to journalArticle

Mochida, K, Hasegawa, A, Li, MW, Fray, MD, Kito, S, Vallelunga, JM, Lloyd, KCK, Yoshiki, A, Obata, Y & Ogura, A 2013, 'High Osmolality Vitrification: A New Method for the Simple and Temperature-Permissive Cryopreservation of Mouse Embryos', PLoS One, vol. 8, no. 1, e49316. https://doi.org/10.1371/journal.pone.0049316
Mochida, Keiji ; Hasegawa, Ayumi ; Li, Ming Wen ; Fray, Martin D. ; Kito, Seiji ; Vallelunga, Jadine M. ; Lloyd, Kevin C K ; Yoshiki, Atsushi ; Obata, Yuichi ; Ogura, Atsuo. / High Osmolality Vitrification : A New Method for the Simple and Temperature-Permissive Cryopreservation of Mouse Embryos. In: PLoS One. 2013 ; Vol. 8, No. 1.
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