Production of transgenic mice from vitrified pronuclear-stage embryos

H. Bagis, H. Odaman, H. Sagirkaya, A. Dinnyés

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Cryopreservation of pronuclear-stage embryos would be useful for transgenic technology and genome preservation purposes. We compared a novel vitrification technique (solid surface vitrification, SSV) with another vitrification method in straws for cryosurvival and to generate transgenic progeny from cryopreserved mouse zygotes following microinjection. The SSV solution consisted of 35% ethylene glycol (EG), 5% polyvinyl-pyrrolidone (PVP), and 0.4 M trehalose in M2 supplemented with 4 mg/ml BSA; the in straw vitrification solution was 7 M EG in M2 plus BSA. In experiment I, we compared the effect of the vitrification solutions alone, without cooling. No reduction was detected in survival and cleavage rates. In experiment II, SSV yielded a significantly higher percentage of morphologically normal zygotes (96%) that also cleaved at significantly higher rates (80%) when compared to that following "in straw" vitrification (68 and 66%, respectively). Cleavage rate in the non-vitrified control group (93%) was significantly higher than that of both vitrified groups. Following embryo transfer, there was no difference in the rate of pups obtained from the SSV, "in straw" vitrified, and control groups (97/ 457, 21%; 15/75, 20% and 56/209, 27%, respectively). In experiment III, SSV vitrified and fresh embryos were used for pronuclear DNA injection. Survival rate of vitrified embryos after microinjection was reduced compared to nonvitrified ones (64 vs. 72%, respectively; P <0.05); however, development to two-cell stage was not different (76 vs. 72%, respectively). Following embryo transfer of vitrified vs. fresh microinjected embryos, in both cases 10% live pups were generated, including transgenic pups. The results demonstrated that the efficiency of generating transgenic pups from SSV vitrified pronuclear zygotes is comparable to that from fresh embryos.

Original languageEnglish
Pages (from-to)173-179
Number of pages7
JournalMolecular Reproduction and Development
Volume61
Issue number2
DOIs
Publication statusPublished - 2002

Fingerprint

Vitrification
Transgenic Mice
Embryonic Structures
Zygote
Ethylene Glycol
Embryo Transfer
Microinjections
Polyvinyls
Pyrrolidinones
Control Groups
Trehalose
Cryopreservation

Keywords

  • Embryo
  • Microinjection
  • Mouse
  • SSV
  • Vitrification

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

Production of transgenic mice from vitrified pronuclear-stage embryos. / Bagis, H.; Odaman, H.; Sagirkaya, H.; Dinnyés, A.

In: Molecular Reproduction and Development, Vol. 61, No. 2, 2002, p. 173-179.

Research output: Contribution to journalArticle

Bagis, H. ; Odaman, H. ; Sagirkaya, H. ; Dinnyés, A. / Production of transgenic mice from vitrified pronuclear-stage embryos. In: Molecular Reproduction and Development. 2002 ; Vol. 61, No. 2. pp. 173-179.
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