Influence Of Adding Arabogalactan Or Ficoll In Freezing Liquid On The Refrigerating Effect Of Cattle Biopsied Embryos

Since the introduction of PCR sex identification technology to production,the birth rate of female calf of dairy cow embryo transplantation has been increased greatly.However, the freezing tolerance of the biopsied embryos was reduced due to a certain degree of injury led in the sampling operation. In this study, a selection protocol was conducted to find out several prescriptions of freezing liquid which were more suited to the embryos of which zona pellucida were damaged and the amount of cells reduced through adding the different concentrations of arabinogalactan and ficoll to the conventional refrigerating fluid of embryos based on ethlene glycol.Three experiments were done. Test one: observing test on refreezing-thawing frozen-thawed embryos sampled; Test two: culturing test on refreezing-thawing frozen-thawed embryos sampled; Test three: culturing test on freezing-thawing sampled fresh embryos; Test four:transplanting test on freezing-thawing sampled fresh embryos. Through the three experiments above,0.03EAS1 and 0.6EFS1 were selected respectively from the arabinogalactan series of frozen liquid and the ficoll series of frozen liquid, and then these two frozen liquid and EG FREEZE(imports of ethylene glycol frozen liquid) were used to freeze biopsied embryos and integrity embryos in vitro of bovine.The freezing effect were compared.Results: Test one: the embryo recoveries、0.5h morphologically integrity rate、1h morphologically integrity rate、1.5h morphologically integrity rate of all kinds of refrigerating fluid were compared with control group,and then six sorts of refrigerating fluid whose refrigerating effect was better had been screened out from arabogalactan series refrigerating fluid: 0.005EAS1、0.01EAS1、0.02EAS1、0.03EAS1、0.04EAS1、0.05EAS1,the contrast results of the four indicators were: 96.67%、96.67%、93.33%、93.33%、93.33%、96.67%vs90.00%;72.41%、65.52%、57.14%、85.71%、75.00%、65.52%vs61.62%;58.62%、55.17%、42.86%、78.57%、71.43%、48.28%vs51.52%;51.72%、41.38%、39.29%、60.71%、67.86%、37.93%vs39.39%.Seven sorts of refrigerating fluid were selected from ficoll series refrigerating fluid: 0.4EFS、0.5EFS、0.6EFS、0.1EFS1、0.2EFS1、0.5EFS1、0.6EFS1,the contrast results of the four indicators were: 90.00%、96.77%、90.63%、100.00%、90.00%、80.65%、100.00%vs90.00%;77.78%、56.67%、65.52%、 70.00%、88.89%、64.00%、76.67% vs61.62%;70.37%、53.33%、55.17%、56.67%、66.67%、52.00%、60.00%vs51.52%;44.44%、46.67%、41.38%、40.00%、51.85%、40.00%、60.00%vs39.39%.Test two: the embryo recoveries、developmental rate after culturing 24h、developmental rate after culturing 48h of all kinds of refrigerating fluid were compared with control group,and then 0.03EAS1 and 0.04EAS1 were futher screened out from arabogalactan series refrigerating fluid, the contrast results of the three indicators were: 100.00%、70.00%vs75.00%;20.00%、42.86%vs13.33%;30.00%、42.86%vs13.33%. 0.5EFS and 0.6EFS1were futher screened out from ficoll series refrigerating fluid, the contrast results of the three indicators were: 100.00%、36.36%vs75.00%;35.00%、37.50%vs13.33%;40.00%、37.50%vs13.33%. Test three: the embryo recoveries、developmental rate after culturing 24h、developmental rate after culturing 48h of all kinds of refrigerating fluid were compared with control group,and then the final result was derived: 0.03EAS1(86.05%vs93.62%、70.27%vs15.91%、81.08%vs22.73%)was the best refrigerating fluid in arabogalactan series refrigerating fluid,and 0.6EFS1(88.10%vs93.62%、64.86%vs15.91%、70.27%vs22.73%)was the best refrigerating fluid in ficoll series refrigerating fluid. Transplanting results of test four are consistent with the above. Culturing test on freezing-thawing fresh embryo in vitro: the embryo recoveries、4h morphologically integrity rate、developmental rate after culturing 24h of the three kinds of refrigerating fluid were compared with each other,and then the result was derived: 0.03EAS(190.00%、55.56%、11.11%)was the best refrigerating fluid for biopsied embryos and EG FREEZE(100.00%、55.56%、22.22%)was the best refrigerating fluid for integrity embryos.Conclusion: Better refrigerating effect was acquisitived after adding proper quantities of arabogalactan or ficoll to the routine refrigerating fluid of biopsied embryos,and arabogalactan was better than ficoll. For integrity embryos, better refrigerating effect was acquisitived from EG FREEZE.