| Programmed freezing and vitrification freezing respectively implemented in stage 4 cells in mice,8 cells,and 3-4 blastocysts horizontal comparative studyObjective: this experiment with 4,8 cells in mice and blastocyst embryo as experimental object,with Cryoloop as the carrier,separately in two different ways of frozen,discuss and compare the two kinds of freezing method in different periods of mice embryo freezing effect.Methods:1 Kunming mice,in vitro fertilization(ivf),in 48 ~ 50 hours after HCG,60 to 62 hours and 4 cells from 85 ~ 96 hours,8 cell and a period of 3 to 4 blastocyst vitrification freezing and stylized.2 The glass transition of frozen and recovery: embryo first with a balanced liquid 7.5%(v/v)EG,7.5%(v/v)DMSO balance 2 minutes;And then transferred to 15%(v/v)EG,15%(v/v)DMSO,0.65 M sucrose vitrification freezing liquid,time control within 30 seconds.Embryo recovery by 0.33 M,0.2 M,0 M sucrose recovery liquid,gradient dilution elution cryoprotectants,balance time of 2 min,3 min respectively and 5 min.3 Programmed freezing and recovery: the embryo in the coolant into the straw after processing,into frozen,starting temperature 20 ?,2 ?/ min speed down to-7 ?,keep the 5 minutes-7 ?,artificial planting ice,keep for 10 minutes.At 0.3 ?/ min speed down to-35 ?,-50 ?/ min speed down to-150 ?,from the application in the liquid nitrogen temperature drop out of a straw,in liquid nitrogen in box,putting the straw into a straw into the embryo liquid nitrogen.Results:1 4 cell groups: programmed frozen embryos,104 thawed for 97,the survival rate 93.2%(97/104),to continue to develop after 72 hours,the blastocyst formation rate was 84.5%(82/97),blastocyst hatched rate was 65.9%(54/82).Vitrification of frozen embryos,118 thawed live 109,survival rate 92.3%(109/118),embryo to continue to develop after 72 hours,the blastocyst formation rate was 84.5(82/97),blastocyst hatched rate was 65.9%(54/82).Two groups of 4 cell embryos in survival rate,the rate of blastocyst formation rate and blastocyst hatched no statistically significant difference(P > 0.05)2 8 cell groups: programmed freezing the 110 thawed embryos survived 90,survival rate of 81.8%,and after 72 hours,the blastocyst formation rate was 84.4%(76/90),the blastocyst hatched rate was 76.3%(58/76).Vitrification of frozen embryos,112 thawed for 102,91.1% survival rate,embryo culture to 72 hours after the blastocyst formation rate was 93.2%(97/112),the blastocyst hatched rate was 88.7%(86/97).Two groups of 8 cell embryos in survival rate,blastocyst formation rate and blastocyst hatched rate difference was statistically significant3 Blastocyst groups programmed frozen embryos,101 thawed live 83,survival rate 82.2%(83/101),after freezing and thawing blastocyst hatched rate was 61.4%(51/83).Vitrification of frozen embryos,92 thawed for 85,92.4% survival rate.After freezing and thawing blastocyst hatched rate was 82.4%(70/85).Two groups of blastocyst respectively for vitrification freezing and programmed freezing and thawing in survival rate,blastocyst hatched rate difference was statistically significantConclusion: in the 4 cell stage,8 cell stage and blastula stage embryos in two groups each respectively for vitrification freezing and programmed freezing,Vitrification freezing treatment 4 cell embryos and programmed frozen embryos in recovery rate,blastocyst formation rate and blastocyst hatched rate has no difference.Vitrification freezing treatment of 8 cell embryos in recovery rate,blastocyst formation rate and blastocyst hatched rate were higher than 8 cell embryos programmed freezing.Blastocyst vitrification freezing processing on recovery rate and hatching rate were higher than programmed freezing blastocysts.Vitrification freezing technology than programmed freezing technology in the frozen 8 and blastula embryonic cells have more advantages... |
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