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A New selection criterion to assess good quality ovine blastocysts after vitrification and to predict their transfer into recipients

Leoni, Giovanni Giuseppe and Berlinguer, Fiammetta and Succu, Sara and Bebbere, Daniela and Mossa, Francesca and Madeddu, Manuela and Ledda, Sergio and Bogliolo, Luisa and Naitana, Salvatore (2008) A New selection criterion to assess good quality ovine blastocysts after vitrification and to predict their transfer into recipients. Molecular Reproduction and Development, Vol. 75 (2), p. 373-382. eISSN 1098-2795. Article.

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DOI: 10.1002/mrd.20754

Abstract

The feasibility to accurately select viable embryos would be valuable for improving pregnancy rates and avoiding futile transfer attempts. The aim of our study was to assess if in vitro-produced embryo quality could be determined by the timing of blastocoelic cavity re-expansion after vitrification, warming, and in vitro culture using sheep as a model. Blastocysts were produced in vitro, vitrified/warmed, and cultured in TCM-199 plus 10% FCS for 72 hr. Embryos were divided into two groups: re-expanded within 8 hr (A) and from 8 to 16 hr (B) of IVC after warming. Fast re-expanded blastocysts showed higher in vitro hatching rates and total cell number calculated on the hatched blastocysts compared with slow re-expanded ones (P < 0.01). Peroxide status evaluation (P < 0.01) and TUNEL test (P < 0.05) revealed a higher number of positive cells in group B compared with group A. The quantitative analysis of protein synthesis revealed a higher synthesis in fast compared with slow re-expanded embryos (P < 0.05). Quantitative RT-PCR showed that 90-kDa Heat Shock Protein was more expressed in group A than in group B (P < 0.05), while the quantity of P34cdc2, Cyclin b, Aquaporin 3, Na/K ATPase, and Actin did not differ between the two groups. Pregnancy rates after transfer to synchronized recipients were higher in fast compared to slow re-expanded blastocysts (P < 0.05). Our results evidenced that timing of blastocoelic cavity re-expansion after vitrification/warming and in vitro culture can be considered as a reliable index of in vitro produced embryo quality and developmental potential.

Item Type:Article
ID Code:1702
Status:Published
Refereed:Yes
Uncontrolled Keywords:Re-expansion time, developmental potential, DNA fragmentation, gene expression, protein synthesis
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/10 Clinica ostetrica e ginecologia veterinaria
Area 07 - Scienze agrarie e veterinarie > VET/02 Fisiologia veterinaria
Area 05 - Scienze biologiche > BIO/10 Biochimica
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze fisiologiche, biochimiche e cellulari
001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
001 Università di Sassari > 01 Dipartimenti > Biologia animale
Publisher:Wiley-Liss
eISSN:1098-2795
Additional Information:Pubblicato online il 17 Aprile 2007.
Deposited On:18 Aug 2009 10:05

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