Vitrification of porcine embryos at various developmental stages using different ultra-rapid cooling procedures

Cuello, C.; Gil, M.A.; Parrilla, Inmaculada; Tornel, Jose; Vázquez, Juan M.; Roca, Jordi; Berthelot, F.; Martinat-Botté, F.; Martı́nez, Emilio A.

doi:10.1016/j.theriogenology.2003.10.007

Cited by 64 publications

(37 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Typical experimental matrices are given in Table 3 for the open pulled straw and the quartz microcapillary. As shown in Figure 5, the threshold CPA concentration associated with the quartz microcapillary is at least 2 M less than that of the traditional straw and 1 M less than the open pulled straw (or the so-called superfine open pulled straw, which is not different from those of the open pulled straw, in agreement with the results reported in reference [8]). It is also shown that 1,2-propanediol (PROH) is superior to ethylene glycol (EG) in terms of the capability to achieve apparent vitrification.…”

Section: Apparent Vitrification Of Cpa Laden Solutionssupporting

confidence: 90%

“…The capillary tubes used include the traditional French type straw (TS, Wipak Medical, Germany), the open pulled straw (OPS, Wipak Medical, Germany) and the thin walled quartz micro-capillary (QMC, Wolfgang Muller Glas Technik, Germany) as shown in Figure 1. Also shown in this figure is the so called superfine open pulled straw (SOPS, Wipak Medical, Germany) which is not significantly different from the open pulled straw either visually or in terms of vitrification capability [8]. A summary of the dimensions of the above-mentioned devices is given in Table 1.…”

Section: Vitrification Devicesmentioning

confidence: 99%

See 1 more Smart Citation

Vitrification by ultra-fast cooling at a low concentration of cryoprotectants in a quartz micro-capillary: A study using murine embryonic stem cells

et al. 2008

View full text Add to dashboard Cite

Conventional cryopreservation protocols for slow-freezing or vitrification involve cell injury due to ice formation/cell dehydration or toxicity of high cryoprotectant (CPA) concentrations, respectively. In this study, we developed a novel cryopreservation technique to achieve ultra-fast cooling rates using a quartz microcapillary (QMC). The QMC enabled vitrification of murine embryonic stem (ES) cells using an intracellular cryoprotectant concentration in the range used for slowing freezing (1-2 M). The cryoprotectants used included 2 M 1,2-propanediol (PROH, cell membrane permeable) and 0.5 M extracellular trehalose (cell membrane impermeable). More than 70% of the murine ES cells post-vitrification attached with respect to non-frozen control cells, and the proliferation rates of the two groups were similar. Preservation of undifferentiated properties of the pluripotent murine ES cells post vitrification cryopreservation was verified using three different types of assays: the expression of transcription factor Oct-4, the presentation of the membrane surface glycoprotein SSEA-1, and the elevated expression of the intracellular enzyme alkaline phosphatase. These results indicate that vitrification at a low concentration (2 M) of intracellular cryoprotectants is a viable and effective approach for the cryopreservation of murine embryonic stem cells.

show abstract

Section: Apparent Vitrification Of Cpa Laden Solutionssupporting

confidence: 90%

Section: Vitrification Devicesmentioning

confidence: 99%

Vitrification by ultra-fast cooling at a low concentration of cryoprotectants in a quartz micro-capillary: A study using murine embryonic stem cells

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Pig blastocysts at later stages are known to be more cryotolerant than early cleavage stage embryos [45][46][47][48]. In this study, however, morulae and early blastocysts were vitrified, and the viability of the embryos was similar to or higher than in previous studies [49,50], indicating the effectiveness of the HFV method.…”

Section: Discussionsupporting

confidence: 39%

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

View full text Add to dashboard Cite

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...

show abstract

“…The success of vitrification seems to be determined by the development stage of a pig embryo since this affects its susceptibility to chilling damage (Nagashima et al 1992;Dobrinsky and Johnson 1994;Leibo et al 1996;Cuello et al 2004;Sánchez-Osorio et al 2008). Cryopreserving embryos at the blastocyst stage will improve embryo selection.…”

Section: Discussionmentioning

confidence: 99%

Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification

Morató

Castillo‐Martín

Yeste

et al. 2016

Reprod. Fertil. Dev.

View full text Add to dashboard Cite

The aim of our study was to assess whether the cryotolerance of in vitro-produced embryos could be influenced by the length of in vitro culture and size of blastocoel cavity before vitrification, using the pig as a model. For this purpose we analysed the cryoresistance and apoptosis rate of blastocysts at different stages of development as derived on Day 5 and 6 of in vitro culture. Blastocysts were subsequently vitrified, warmed and cultured for 24h. Re-expansion rates were recorded at 3 and 24h and total cell number and apoptotic cells were determined at 24h. Day-6 blastocysts showed the highest rates of survival after warming, which indicates higher quality compared with Day-5 blastocysts. Higher re-expansion rates were observed for expanded blastocysts and those in the process of hatching when compared with early blastocysts. Total cell number and apoptotic cells were affected by blastocyst stage, vitrification-warming procedures and length of in vitro culture, as expanding and hatching-hatched blastocysts from Day 6 presented higher percentages of apoptotic cells than fresh blastocysts and blastocysts vitrified at Day 5. Our findings suggest that the cryotop vitrification method is useful for the cryopreservation of porcine blastocysts presenting a high degree of expansion, particularly when vitrification is performed after 6 days of in vitro culture. Furthermore, these results show that faster embryo development underlies higher blastocyst cryotolerance and provide evidence that blastocoel cavity expansion before vitrification is a reliable index of in vitro-produced embryo quality and developmental potential.

show abstract

Vitrification of porcine embryos at various developmental stages using different ultra-rapid cooling procedures

Cited by 64 publications

References 21 publications

Vitrification by ultra-fast cooling at a low concentration of cryoprotectants in a quartz micro-capillary: A study using murine embryonic stem cells

Vitrification by ultra-fast cooling at a low concentration of cryoprotectants in a quartz micro-capillary: A study using murine embryonic stem cells

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

Cryotolerance of porcine in vitro-produced blastocysts relies on blastocyst stage and length of in vitro culture prior to vitrification

Contact Info

Product

Resources

About