Thermal and clinical performance of a closed device designed for human oocyte vitrification based on the optimization of the warming rate

Gallardo, Miguel; Hebles, María; Migueles, Beatriz; Dorado, M.; Aguilera, L.; González, Mercedes Gómez; Piqueras, Paloma; Montero, Lorena; Sánchez-Martín, Pascual; Sánchez-Martín, Fernando; Risco, Ramón

doi:10.1016/j.cryobiol.2016.06.001

Cited by 10 publications

(4 citation statements)

References 34 publications

(53 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Relevant parameters that determine the glass-forming tendency of the cytosol, such as the viscosity of the milieu and the degree of hydrogen bonding between present water and intracellular macromolecules, could for this reason differ between oocytes prepared to vitrification with each protocol 22,23 . It should also be considered that we employed a carrier that is reported to produce higher rates of cooling and warming than others 15,24–30 which may compensate for a possible lower intracellular glass forming tendency in the cells after DP 9,31 .…”

Section: Discussionmentioning

confidence: 99%

“…Note that the loading, sealing and cooling of the vitrification device were performed during the 60 seconds of exposure to VS, and not afterwards. The process of loading the vitrification carrier is described thoroughly elsewhere 24 .…”

Section: Methodsmentioning

confidence: 99%

“…After 60 seconds, oocytes were transferred to a 0.5M sucrose solution for three minutes and eventually placed for 5 minutes in a CPA-free solution. According to previous published measurements employing the same closed vitrification carrier, the zygotes were subjected to warming rates of 200.000 °C/min 24 .…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Human oocytes and zygotes are ready for ultra-fast vitrification after 2 minutes of exposure to standard CPA solutions

Gallardo

Saenz

Risco

2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

Vitrification of human oocytes and embryos in different stages of development is a key element of daily clinical practice of in vitro fertilization treatments. Despite the cooling and warming of the cells is ultra-fast, the procedure as a whole is time consuming. Most of the duration is employed in a long (8–15 minutes), gradual or direct exposure to a non-vitrifying cryoprotectant solution, which is followed by a short exposure to a more concentrated vitrifying solution. A reduction in the duration of the protocols is desirable to improve the workflow in the IVF setting and reduce the time of exposure to suboptimal temperature and osmolarity, as well as potentially toxic cryoprotectants. In this work it is shown that this reduction is feasible. In silico (MatLab program using two-parameter permeability model) and in vitro observations of the oocytes’ osmotic behaviour indicate that the dehydration upon exposure to standard cryoprotectant solutions occurs very fast: the point of minimum volume of the shrink-swell curve is reached within 60 seconds. At that point, intracellular water ejection is complete, which coupled with the permeation of low molecular weight cryoprotectants results in similar intracellular and extracellular solute concentrations. This shows that prolonging the exposure to the cryoprotectant solutions does not improve the cytosolic glass forming tendency and could be avoided. To test this finding, human oocytes and zygotes that were donated for research were subjected to a shortened, dehydration-based protocol, consisting of two consecutive exposures of one-minute to two standard cryoprotectant solutions, containing ethylene glycol, dimethyl sulfoxide and sucrose. At the end of this two-minute dehydration protocol, the critical intracellular solute concentration necessary for successful vitrification was attained, confirmed by the post-warming survival and ability to resume cytokinesis of the cells. Further studies of the developmental competency of oocytes and embryos would be necessary to determine the suitability of this specific dehydration protocol for clinical practice, but based on our results, short times of exposure to increasingly hypertonic solutions could be a more time-efficient strategy to prepare human oocytes and embryos for vitrification.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Human oocytes and zygotes are ready for ultra-fast vitrification after 2 minutes of exposure to standard CPA solutions

Gallardo

Saenz

Risco

2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Indeed, a series of studies by the group of the late Peter Mazur showed that high warming rates are very important in the vitrification of mouse oocytes and embryos, in fact more important than rapid cooling (Mazur and Paredes 2016). Whether this translates to oocytes of other species, including horses, is not clear, but it should be kept in mind during the process (Gallardo et al 2016).…”

Section: Speed Of Vitrification and Warmingmentioning

confidence: 99%

Cryopreservation of equine oocytes: looking into the crystal ball

Coster

Angel‐Velez

Soom

et al. 2020

Reprod. Fertil. Dev.

View full text Add to dashboard Cite

In vitro embryo production has evolved rapidly in the horse over the past decade, but blastocyst rates from vitrified equine oocytes remain quite poor and further research is needed to warrant application. Oocyte vitrification is affected by several technical and biological factors. In the horse, short exposure of immature oocytes to the combination of permeating and non-permeating cryoprotective agents has been associated with the best results so far. High cooling and warming rates are also crucial and can be obtained by using minimal volumes and open cryodevices. Vitrification of in vivo-matured oocytes has yielded better results, but is less practical. The presence of the corona radiata seems to partially protect those factors that are necessary for the construction of the normal spindle and for chromosome alignment, but multiple layers of cumulus cells may impair permeation of cryoprotective agents. In addition to the spindle, the oolemma and mitochondria are also particularly sensitive to vitrification damage, which should be minimised in future vitrification procedures. This review presents promising protocols and novel strategies in equine oocyte vitrification, with a focus on blastocyst development and foal production as most reliable outcome parameters.

show abstract

Safety and efficiency of oocyte vitrification

Munck

Vajta

2017

Cryobiology

View full text Add to dashboard Cite

Thermal and clinical performance of a closed device designed for human oocyte vitrification based on the optimization of the warming rate

Cited by 10 publications

References 34 publications

Human oocytes and zygotes are ready for ultra-fast vitrification after 2 minutes of exposure to standard CPA solutions

Human oocytes and zygotes are ready for ultra-fast vitrification after 2 minutes of exposure to standard CPA solutions

Cryopreservation of equine oocytes: looking into the crystal ball

Safety and efficiency of oocyte vitrification

Contact Info

Product

Resources

About