Transfer of Vitrified Sheep Morula

Abstract

Contents The aim of the experiment was to determine the susceptibility to vitrification of the sheep morula. As a vitrification medium the mixture of 35% glycerol + 30% 1,2 propanediol or 30% glycerol + 35% 1,2 propanediol was used. The embryos were fiozen in glass ampules placed for 5 min in nitrogen uapour before plunging to liquid nitrogen. Dilution after rapid thawing was done in one step in a 1M sucrose solution. The pregnancies were obtained only when the embryos were transferred immediately after thawing. Of 8 recipients which received 15 embryos 6 became pregnant. When the transfer were done 30–60 minutes after thawing, none of 4 recipients which received 8 embryos conceived. Inhalt: Transfer von stungefrorenen Morulae vom Schaf In diesem Versuch sollte die Eignung von Schafsmorulae fur die Sturzgefrierung untersucht werden. Als Vitrifikationsmedium wurde eine Mischung von 35 % Glycerin + 30% Propanediol oder von 30% Glycerin f 35 % 1,2‐Propanediol benutzt. Die Embryonen wurden in Glasampullen eingefioren, die vor dem Eintauchen in jliissigen Stickstoff fur 5 min im Stickstoffdampf gehalten wurden. Nach dem schnellen Auftauen erfolgte die Ausverdiinnung in 1M‐Sucrose‐Losung in einer Stufe. Trachtigkeiten wurden nur erzielt, wenn die Embryonen unmittelbar nach dem Auftauen iibertragen wurden. Von 8 Empfangern, die 15 Embryonen erhielten, wurden 6 tragend. Erfolgten die Transfers erst 30–60 min nach dem Auftauen, dann wurde keiner von 4 Empfangern, in die 8 Embryonen iibertragen wurden, tragend.

Oocyte and embryo cryopreservation – state of art and recent developments in domestic animals

“…First Willadsen et al (1976), then Moor and Bilton (1977) and finally Smorąg et al (1977) obtained live lambs following transfer of sheep morulae and blastocysts. The first successful vitrification of ovine embryos was carried out by Gajda et al (1989). We demonstrated that the survival of transferred vitrified sheep morulae (60%) is comparable to that obtained after transfer of embryos frozen using the conventional slow method.…”

Viabilidad, maduración y desarrollo embrionario in vitro de ovocitos inmaduros de porcino y ovino, vitrificados en diferentes recipientes

“…This may be explained because pig oocytes and embryos are characterized by a high lipid content, which is stored as droplets in the cytoplasm [38,41]. This high lipid content has a negative influence on successful manipulations of oocytes and embryos, thereby resulting in cryopreservation methods with less success for pig than in bovine [19]. This is supported on the low rates of ED reported by Nagashima et al [36] and Wu et al [52] or blastocyst formation reported by Somfai et al [45].…”

“…-La viabilidad observada en los ovocitos inmaduros de porcino y ovino con el proceso de vitrificación ha generado información que parece indicar que los ovocitos de porcino resisten menos al proceso de vitrificación debido al alto contenido de lípidos en su citoplasma (Gajda, 2009). Por consiguiente, se considera que los ovocitos de ovino tendrán un mayor porcentaje de viabilidad, maduración y desarrollo embrionario después de la vitrificación con respecto a los ovocitos porcinos sometidos a las mismas condiciones de criopreservación.…”

“…El desarrollo de la criopreservación se ha traducido en logros tales como la tecnología para el almacenamiento de embriones de mamíferos como vacas, ovejas, cerdos y otras especies (Esaki et al, 2004). Sin embargo, los ovocitos siguen siendo difíciles de ser criopreservados debido a su sensibilidad a la refrigeración y congelación (Gupta et al, 2007;Gajda, 2009). La vitrificación es un procedimiento para la conservación de ovocitos y embriones en el que se disminuye la formación 2 de cristales de hielo, que pueden dañar los componentes morfológicos y funcionales de las células .…”

“…The development of cryopreservation has resulted in achievements such as technology for the storage of mammalian embryos of livestock including cattle, sheep, pigs and other species [16]. Nevertheless, mammalian oocytes remain one of the most difficult cell types to be successfully cryopreserved due to their sensitivity to cooling and freezing [21,19]. Vitrification is a procedure for preserving oocytes and embryos in which there is no formation of ice crystals, which could damage the morphological and functional cell components [49].…”