Abstract
This study was carried out to establish an appropriate condition for the efficient cryopreservation of the mouse pronuclear embryo. in vitro cryopreservation ot pronuclear embryos was carried out by slow Ireezing or vitrification methods and development rate of 2-cell, blastocyst and hatched blastocyst was measured as well as survival rate of the thawed pronuclear embryo. After slow freezing, vitrification and thawing of mouse pronuclear embryos, the survival rate and blastocyst development rate for the vitrification group was 97.3 and 53.4%, respectively, which was significantly higher as compared to the slow freezing group with 88.6 and 23.9%, respectively (p<0.05). Blastocyst developmental rate in each experimental group was significantly higher for 21 h in the post-hCG group at 40.5-57.0% than the 24 h post-hCG group at 40.5% (p<0.05). ICM (Inner cell mass) cell numbers of blastocyst-stage embryos during the different stages of mouse pronuclear embryos, slow freezing and vitrification period in the control and vitrification groups were 22.1±2.7 and 17.0±3.1∼22.0±3.2, respectively; hence, the slow freezing group (10.2±2.0) had significantly higher cell numbers than those of the other two groups (p<0.05). Trophoblast (TE) cell number in the control group, 65.8±12.6, was significantly higher than in the slow freezing group, 41.6±11.1 (p<0.05). The total cell numbers in the control group and 21 h post hCG group were 87.9±13.6 and 81.8+14.1, respectively, and were significantly higher than for the slow freezing group (51.8±12.6; p<0.05).
Highlights
Since the first report on the production of first offspring (Whittngham et al, 1972) from the mammalian cryopreserved embryo transfer, the studies are continuously carrying out to optimize the condition for freezing and thawing and various mammalians are producing offspring from cryopreserved embryo transfer
The samples for control and experimental groups were taken after sorting of pronuclear embryos at 21 h post hCG injection
After the study on cryopreservation of mouse embryos by Whittingham et al (1972), many researchers used a variety of species embryos to study the components of cryoprotectant and freezing method for optimization of freezing condition and to increase the survival rate of embryos after cryopreservation
Summary
Since the first report on the production of first offspring (Whittngham et al, 1972) from the mammalian cryopreserved embryo transfer, the studies are continuously carrying out to optimize the condition for freezing and thawing and various mammalians are producing offspring from cryopreserved embryo transfer. The collected embryos were washed would be able to produce a decrease in embryo three times with culture media and treated with culture developmental rate when they are thawed and fertilized. The cortical granules are observed in Metaphase II stage during selected normal pronuclear embryos were cultured with G1 the exposure of cryoprotectant and freezing (Stachecki et al, (G1-5, Vitrolife, Sweden), culture medium 37°C. Various survival and developmental rates are according to the experimental conditions, further, they were produced according to oocyte developmental stages cultured with control group after slow freezing and (Mandelbaum et al, 1998). The embryos were embryo cryopreservation with an investigation of exposed to media containing 1.5 M PROH for 10 min, and survivability, embryo developmental rate and ICM
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