A successful and easy method for equine embryo vitrification is currently being used for commercial purposes. Anyway, cryopreservation is more successful when embryos are less than 300 µm, day 6 after ovulation, but in commercial programs embryos are often collected on day 7 or 8, for an higher recovery rate. Recently, successful cryopreservation of equine expanded blastocysts after blastocoele collapse was obtained using a micromanipulator. The aim of this study was to test a field method to simplify collapse and cryopreservation procedures for expanded equine blastocysts. Equine blastocysts > 400 µm were collected from 8 mares 7-9 days after ovulation and cut with a 27 gauge syringe needle before standard in straw vitrification. Embryos were warmed and transferred in 6 recipients or stained with Hoechst 33342/propidium iodide. 13 embryos were collected from 20 flushes, and none of 6 produced pregnancy after warming and transfer. Only 1/7 embryo was intact after warming and had about 20% dead cells. Use of standard vitrification and solutions seems to be inadequate to assure large embryos survival. The method for blastocoele collapse used in this study should be improved to enhance fluid loss and tested with a non-standard vitrification protocol, to establish its efficacy.
Merlo B, Mislei B, Bulfone G, Rizzato G, Iacono E, Mari G. (2013). Vitrification of equine expanded blastocysts. [10.4488/SIRA.2013.X].
Vitrification of equine expanded blastocysts.
MERLO, BARBARA;MISLEI, BEATRICE;RIZZATO, GIOVANNI;IACONO, ELEONORA;MARI, GAETANO
2013
Abstract
A successful and easy method for equine embryo vitrification is currently being used for commercial purposes. Anyway, cryopreservation is more successful when embryos are less than 300 µm, day 6 after ovulation, but in commercial programs embryos are often collected on day 7 or 8, for an higher recovery rate. Recently, successful cryopreservation of equine expanded blastocysts after blastocoele collapse was obtained using a micromanipulator. The aim of this study was to test a field method to simplify collapse and cryopreservation procedures for expanded equine blastocysts. Equine blastocysts > 400 µm were collected from 8 mares 7-9 days after ovulation and cut with a 27 gauge syringe needle before standard in straw vitrification. Embryos were warmed and transferred in 6 recipients or stained with Hoechst 33342/propidium iodide. 13 embryos were collected from 20 flushes, and none of 6 produced pregnancy after warming and transfer. Only 1/7 embryo was intact after warming and had about 20% dead cells. Use of standard vitrification and solutions seems to be inadequate to assure large embryos survival. The method for blastocoele collapse used in this study should be improved to enhance fluid loss and tested with a non-standard vitrification protocol, to establish its efficacy.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.