Birth of piglets after OPS vitrification and transfer of compacted morula stage embryos with intact zona pellucida

Berthelot, F.; Martinat-Botté, F.; Perreau, C.; Terqui, M.

doi:10.1051/rnd:2001129

Cited by 70 publications

(40 citation statements)

References 14 publications

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“…Vajta [3] found that reducing this amount leads to faster cooling and a warming rate that allows embryos to rapidly pass through particular critical temperature zones, so that a higher survival rate was obtained compared to conventional vitrification. In fact, piglets have been successfully produced after transferring ultrarapidly vitrified embryos to recipient pigs [5][6][7][8]. The cooling and warming rate for the MVC-method used here is considered higher than that for conventional vitrification, because (i) the volume of vitrification medium surrounding embryos is smaller due to the excess solution being removed by pipetting after embryos were placed on the Cryotop blade and (ii) the embryos placed on the MVC plate can be directly exposed to LN and the warming medium in contrast to embryos being covered by a tip of a container such as GLT or ST filled with vitrification solution [12,13].…”

Section: Discussionmentioning

confidence: 99%

“…T h i s t e c h n i q u e i s a s t a n d a r d cryopreservation procedure for porcine embryos mainly at the blastocyst stage [4][5][6][7][8]. In fact, several studies have reported live offspring production after transfer of vitrified porcine embryos [5][6][7][8].…”

mentioning

confidence: 99%

“…In fact, several studies have reported live offspring production after transfer of vitrified porcine embryos [5][6][7][8]. On the other hand, however, the pregnancies and numbers of piglets produced from vitrified embryos have been extremely low and should be improved by optimizing vitrification procedures for practical use.…”

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confidence: 99%

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Improved Survival of Vitrified In Vivo-Derived Porcine Embryos

Ushijima

Yoshioka

Esaki

et al. 2004

Journal of Reproduction and Development

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Abstract. An efficient cryopreservation protocol for porcine morulae was investigated with three types of vitrification having different cooling rates (Exp. 1). Survival of embryos vitrified after removal of cytoplasmic lipid droplets was also examined by means of the minimum volume cooling (MVC) method (Exp. 2). In Exp. 1, the morula stage embryos were vitrified with a 0.25 ml plastic straw (ST-method), gel loading tip (GLT-method) and the MVC-method, respectively, and stored in liquid nitrogen after which they were warmed in sucrose solutions with cryoprotectants being subsequently removed in a stepwise manner. In Exp. 2, morulae were centrifuged with 7.5 µg/ml cytocharasin B at 12000 × g for 20 min to polarize the cytoplasmic lipid droplets that were then removed from the embryos by micromanipulation (delipation). Both those delipated at the morula stage and the intact embryos at the morula to blastocyst stages were vitrified by the MVC-method. In vitro survival of the vitrified embryos was assessed in both experiments by culturing in NCSU-23 + 10% FCS for 48 h. In vitro developments of vitrified embryos after warming to blastocysts were 20% (6/30) for the ST-method, 39% (18/46) for the GLT-method, and 60% (26/43) for the MVC-method. Embryo survival was further improved by vitrification after delipation (95%, 35/37) compared to intact vitrified morulae (24/42, 57%, P<0.001) and blastocysts (23/31, 74%, P<0.05). Moreover, the number of cells in blastocysts (92 ± 25) derived from the delipated-vitrified morulae was comparable to those derived from intact control non-vitrified embryos (103 ± 31). Our results demonstrate that vitrified porcine morulae have the highest survival when using the MVC-method in conjunction with delipation.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Improved Survival of Vitrified In Vivo-Derived Porcine Embryos

Ushijima

Yoshioka

Esaki

et al. 2004

Journal of Reproduction and Development

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“…Furthermore, the in vitro development of vitrified embryos after rewarming was nearly 100%. Therefore, it is likely that similar numbers of piglets could be obtained by transferring fewer embryos than in this study; in other words, a higher production efficiency of piglets could be achieved.Production of pigs from vitrified in vivo-derived embryos has been achieved using blastocysts [40][41][42][43][44]. Pig blastocysts at later stages are known to be more cryotolerant than early cleavage stage embryos [45][46][47][48].…”

mentioning

confidence: 99%

Hollow Fiber Vitrification: A Novel Method for Vitrifying Multiple Embryos in a Single Device

Matsunari

Maehara

Nakano

et al. 2012

Journal of Reproduction and Development

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Abstract. Current embryo vitrification methods with proven efficacy are based on the minimum volume cooling (MVC) concept by which embryos are vitrified and rewarmed ultrarapidly in a very small amount of cryopreserving solution to ensure the high viability of the embryos. However, these methods are not suitable for simultaneously vitrifying a large number of embryos. Here, we describe a novel vitrification method based on use of a hollow fiber device, which can easily hold as many as 40 mouse or 20 porcine embryos in less than 0.1 µl of solution. Survival rates of up to 100% were obtained for mouse embryos vitrified in the presence of 15% DMSO, 15% ethylene glycol and 0.5 M sucrose using the hollow fiber vitrification (HFV) method, regardless of the developmental stage of the embryos (1-cell, 2-cell, morula or blastocyst; n = 50/ group). The HFV method was also proven to be effective for vitrifying porcine in vitro-and in vivo-derived embryos that are known to be highly cryosensitive. For porcine embryos, the blastocyst formation rate of in vitro maturation (IVM)-derived parthenogenetic morulae after vitrification (48/65, 73.8%) did not decrease significantly compared with non-vitrified embryos (59/65, 90.8%). Transfer of 72 in vivo-derived embryos vitrified at the morula/early blastocyst stages to 3 recipients gave rise to 29 (40.3%) piglets. These data demonstrate that the HFV method enables simultaneous vitrification of multiple embryos while still adhering to the MVC concept, and this new method is very effective for cryopreserving embryos of mice and pigs. Key words: Cryopreservation, Hollow fiber, Mouse embryos, Pig embryos, Vitrification (J. Reprod. Dev. 58: [599][600][601][602][603][604][605][606][607][608] 2012) E mbryo cryopreservation is widely and routinely used both for research in reproductive biology and for the development of practical applications in animal industries and human reproductive medicine. The cryopreservation of oocytes and embryos is essential for the long-term preservation of valuable genetic resources in experimental and livestock animals [1][2][3][4][5], and the cryopreservation of embryos improves pregnancy rates in assisted reproductive technology applications [6][7][8]. Various protocols have been developed for the cryopreservation of animal embryos, and many are used in research and clinical applications [1,5,9].In recent years, embryo vitrification has been applied to an increasing number of animal species, and high post-cryopreservation embryo viability has been achieved [10,11]. The basic concept of vitrification is as follows: a solution containing a high concentration of a cryoprotective agent (CPA) is rapidly cooled, causing it to transform from the liquid phase to the solid phase without forming ice crystals. As a result, cells suspended in the solution are preserved in an ultra-low temperature glassy (amorphous) material [10,12]. To create this glassy state, a solution containing a high concentration of CPA (normally 4-6 M) needs to be cooled ultrarapidly. Howeve...

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“…Transfer of 147 vitrified expanded blastocysts into 10 recipients, resulted in birth of 29 live piglets, with pregnancy rate of 50% (Figure 2; Gajda et al, 2004). Recent application of the OPS method to pig blastocysts has allowed excellent in vitro survival (Berthelot et al, 2000(Berthelot et al, , 2001Gajda and Smorąg, 2001), but pregnancy rates after transfer are quite variable, ranging from zero to 60% (Gajda et al, 2006). …”

Section: Speciesmentioning

confidence: 99%

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

Gajda¹,

Smorąg²

2009

J. Anim. Feed Sci.

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During the past few decades, significant progress in cryopreservation of mammalian oocytes and embryos has been observed. Transfer of cryopreserved embryos or oocytes resulted in live offspring in at least 25 species. So far, two major methods have been used for oocyte and embryo cryopreservation: conventional slow-rate freezing and vitrification. This review summarizes the progress in cryopreservation of mammalian oocytes and embryos that has been achieved by modifying oocyte/embryo susceptibility to cryopreservation or vitrification technology through such techniques as solid-surface vitrification, cryoloop, microdrop, cryotop, electron microscopy grids, nylon mesh, open pulled straw method or removal of lipids, addition of antifreeze protein or cytoskeletonstabilizing agents, cholesterol or liposomes, centrifugation prior to cryopreservation or application of high hydrostatic pressure. In conclusion, the vitrification method opened new perspectives in cryopreservation of embryos and oocytes, both for in vitro fertilized and somatic nuclear transfer. Authors believe that new cryopreservation procedures which modify the susceptibility of oocytes and embryos to cryopreservation will gain importance as a major tool in mammalian gamete and embryo cryopreservation.

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Birth of piglets after OPS vitrification and transfer of compacted morula stage embryos with intact zona pellucida

Cited by 70 publications

References 14 publications

Improved Survival of Vitrified In Vivo-Derived Porcine Embryos

Improved Survival of Vitrified In Vivo-Derived Porcine Embryos

Hollow Fiber Vitrification: A Novel Method for Vitrifying Multiple Embryos in a Single Device

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

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