**9. References**

130 Current Frontiers in Cryopreservation

A clear heat release peak is present during cooling as well as melting during rewarming. (b)

rewarming. (c) Thermal history for OPS when filled with a 1.5M propane-1,2-diol and 0.3 M sucrose cryoprotectant solution quenched in Slush nitrogen and then thawed in a water bath at 37ºC. In this case, crystallization during cooling and melting during rewarming was not recorded. However, visual inspection reveals the presence of ice. (d) Thermal history for QC

quenched in Slush nitrogen and then thawed in a water bath at 37ºC. The sample keeps its transparency over all the cooling–rewarming cycle, an indication of the capability of this

All these changes have allowed us to maintain a concentration of cryoprotectors typical of slow freezing, 2 M PrOH+0.5 M sucrose, obtaining a morphological survival rate of 92 % in human oocytes (31). Dr. Ho-Joon Lee et al (69) tested this new technique on mouse ooctyes and they saw that using Ultra-vitrification with low concentrations of cryoprotectors improved the fertilization rate and above the blastulation rate. Only the use of Ultravit device in this technique ensures the non contamination of the sample or cross-contamination

Surv. rate 61 91,8 92.5 92

Fert. rate 61,3 67,9 75 ?

rate 12 33.1 59.1 ?

lower the concentration of the cryoprotectants decreasing the toxicity in cells.

Table 4. Comparison between slow Freezing, Vitrification and Ultravitrification (78, 69, 31)

This comparison demonstrates the use of low concentration of cryoprotectant in the Ultravitrification protocol favours the morphological survival (92%) and increases the blastulation rate (59.1%). Thus confirming the hypothesis that cryoprotectants are toxic to the biological sample and if we could find a vitrification protocol that would allow us to vitrify without cryoprotectant, we would achieve a better embryo development and a greater chance of pregnancy in the case of freezing eggs or embryos. A lot more studying is needed regarding this new technique but a priori the results indicate that we can hopefully

Cryopreservation protocols have improved and resulted in a much higher efficiency in outcomes in the last years. However, it remains important to always seek for amelioration

Ultravitrification mourine oocytes (69)

Ultravitrification human oocytes (31)

Thermal history for QC when filled with a 1.5M propane-1,2-diol and 0.3M sucrose cryoprotectant solution quenched in LN2 and then thawed in a water bath at 37ºC. Crystallization of water is not obvious during cooling, but melting is shown during

when filled with a 1.5Mpropane-1,2-diol and 0.3 Msucrose cryoprotectant solution

Freezing (78) Vitrification (78)

approach to vitrify the studied solution.

in communal containers.

Blast.

**7. Conclusion** 

% Slow


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**Part 3** 

**Farm / Pet / Laboratory Animal ART** 

