| people are still puzlled by the low of the fertilization rate,cleavage rate,blastocyst rate and clinic pregnancy in the field of assiste reproduction technology. Chemical pollution of the IVF laboratory is considered as the important reason. Chemical pollution mainly comes from discharge of the volatile organic compounds(VOCs).For the IVF laboratory,they come from equipments, painting, cement,floor and laminar flow. When we built the IVF laboratory,we use the chemical engineering materials at the least. In order to test the envriomental quality of the IVF laboratory,we carry out a series of experiments about mouse embryo, including the experiments of ovarian stimulation,occurring of the gamete and embryo in vitro,vitrification for cryopreservation/thawing mouse embryo and assisted hatching . The fertilization, ratecleavage rate,blastocyst rate and clinic pregnancy of the experiment are according with other experiments,even are better than others.At the same time, we found that there are no significant difference between the two results of the 3 months and 1 year after IVF laboratory decoration, the P>0.05. The result point out through reducing the volume of the VOCs in IVF laboratory, the fertilization rate, cleavage rate, blastocyst rate and clinic pregnancy are higher than befor. This is very important for clinical ending.ObjectiveTo test the environment of the new IVF laboratory is suitable for gamete and embryo to culture and development in vitro.Methods1)Femal mouse is injected through intraperitoneal way by Pregnant Mare Serum Gonadotropin (PMSG)10IU/1 mouse(0.1ml), on the first day at 5:00pm, Gonadotropin(HCG)10IU/1 mouse(0.1ml)is injected to femal mouse on the third day. After injection of HCG12-13h, the male mouses are sent to death,obtaining and making sperm capacitation.After injection of HCG13-14h,the femal mouses are sent to death,obainting ovums.Put ovums into the incubator,then put semen which have the capacitation to fertilization, into the cuiture-dish waiting for fertilization.Observing the situation of the fertilization after 4-6h.,then observing cleavage rate,high-quality embryo rate , blastocyst rate for each 24h.2)Obtaining 8-cell embryos: Pregnant Mare Serum Gonadotropin(PMSG)10IU/1 mouse(0.1)is injected to female mouse through intraperitoneal way on the first day 12:00.Gonadotropin(HCG)10IU/1 mouse(0.1)is injected to femal mouse on the third day.Putting femal and male mouses in the same cageon one and one on the same day of HCG .Inspecting bolt on the second morning,bolt picked is considered successful fetilization and mark d0.5. On the sixth day,femal mouses are sent to death ,then getting the embryos,marking d2.5.3) Vitrification: Moving the mouse embryos to vitrification solution1,keeping for about 5-7minutes ,later moving to vitrification solution2,and then loaded into the freezing containing rod,which would be throwed into liquid nitrogen. Adding thaw liquid 1 into 3001 dish,five-well plates is added by unfrozen liquid 2, 3, 4. Rod equipped with embryos is put into the unfrozen liquid 1, staying1 minute, quikly. Followed into the unfrozen liquid 2,3,4,respectively,for3,5,5 minutes.Finally moved into micro-droplets for blastocysts to be observed. Embryos thawed were placed iin HEPES solution. The part of 3 OË‹clock of zona is punched by laser beam at the larger gap around the zona.The embryos were cultured for 5 days after punched,then observing the situation of the embryos.Result1)The total cycles of fertilization in vitro is 10, the total number of the oocytes is 189. The result of the experiment of IVF laboratory decorationed 3 months later is 94 occytes, fertilization rate, cleavage rate, high-quality embryo rate, blastocyst formation rate and blastocyst hatching rate is 84.0%,96.2%,97.3%, 96.1%and 93.1%, respectively. The result of the experiment of IVF laboratory decorationed 1year later is 95 occytes, fertilization rate, cleavage rate, high-quality embryo rate, blastocyst formation rate and blastocyst hatching rate is 87.4%, 92.8%, 98.7%, 97.4%and 93.3%, respectively.2)The result of the experiment of IVF laboratory decorationed 3 months later is 17 embryos frozen, 17 embryos thawed, 16 high-quality embryos, 94.1% high-quality embryo rate, 94.1% blastocyst rate, 100%hatching rate. For the IVF laboratory decorationed 1 year later, the result is 21 embryos frozen, 21 embryos thawed, 21high-quality embryos, 100% high-quality embryo rate, 100% blastocyst rate, 100% hatching rate.Conclusion1)VOCs is the main component of the IVF laboratory's pollution, mainly coming from the decoration materials of the IVF laboratory. There are not obvious differences among fertilization rate, cleavage rate, high quality embryo rate, blastocryst rate and hatching rate, between 3 months and 1 year after IVF laboratory decorated. In further to manifest that the decoration materials with high environmental standards could effectively reduce the VOCs of IVF laboratory. Creating a safe and non-toxic culture environment for gemetes and embryos. It is very important and significant for the outcomes of IVF.2)Vitrification/ thawing and laser-assisted hatching technology is more effective compared with other methods. The methods can get better high quanlity embryo rate after thawed and hatching rate after laser drilling. |
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