Survival of rapidly frozen hatched mouse blastocysts

Csaba Pribenszky, S. Cseh, Zsolt Abonyi-Tóth, L. Solti

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The objective of the present study was to examine the effect of rapid freezing on the in vitro and in vivo survival of zona-pellucida-free hatched mouse blastocysts. Hatched blastocysts were rapidly frozen in a freezing medium containing either ethylene glycol (EG) or glycerol (G) in 1.5 M or 3 M concentration. Prior to freezing, embryos were equilibrated in the freezing medium for 2 min, 10 min, 20 min or 30 min at room temperature. To freeze them, embryos were held in liquid nitrogen vapour [∼1 cm above the surface of the liquid nitrogen (LN2)] for 2 minutes and then immersed into LN 2. After thawing, embryos were transferred either to rehydration medium (DPBS + 10% foetal calf serum + 0.5 M sucrose) for 10 minutes or rehydrated directly in DPBS supplemented with foetal calf serum. In vitro survival of embryos frozen with EG was higher than those frozen with G. The highest survival was obtained with 3 M EG and 2 min or 10 min equilibration prior to freezing, combined with direct rehydration after thawing. Frozen blastocysts developed into normal foetuses as well as unfrozen control ones did, with averages of 30% (control), 26% (EG) and 15% (G). The results show that hatching and hatched mouse blastocysts can be cryopreserved by a simple rapid freezing protocol in EG without significant loss of viability. Our data indicate that the mechanical protection of the zona pellucida is not needed during freezing in these stages.

Original languageEnglish
Pages (from-to)361-366
Number of pages6
JournalZygote
Volume11
Issue number4
DOIs
Publication statusPublished - Nov 2003

Fingerprint

Blastocyst
Freezing
Ethylene Glycol
Embryonic Structures
Zona Pellucida
Fluid Therapy
Nitrogen
Serum
Glycerol
Sucrose
Fetus
Temperature

Keywords

  • Cryopreservation
  • Ethylene glycol
  • Glycerol
  • Hatched blastocyst
  • Mouse embryo

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Survival of rapidly frozen hatched mouse blastocysts. / Pribenszky, Csaba; Cseh, S.; Abonyi-Tóth, Zsolt; Solti, L.

In: Zygote, Vol. 11, No. 4, 11.2003, p. 361-366.

Research output: Contribution to journalArticle

Pribenszky, Csaba ; Cseh, S. ; Abonyi-Tóth, Zsolt ; Solti, L. / Survival of rapidly frozen hatched mouse blastocysts. In: Zygote. 2003 ; Vol. 11, No. 4. pp. 361-366.
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