| In this article we tested the effect of amino acids derived from oviduct fluid(1×OAA) on in vitro development of embryo by in vivo fertilization(1-cell embryo),and the animal was Kunming mouse.The purpose was to seek for an effective OAA composition for embryo in vitro development and optimize culture system.At the same time,it also could be used as a model for other mammalian embryo development.There were four experiments,the conclusions were as follows:(1) At first,we tested the effect of the OAA and sigma's amino acids(SAA) on the development of 1-cell embryo on different stages,while the group of amino acids free was as control.We found that the rate of 2-cell embryo was 88.74%,71.28%and 69.51%respectively,the rate was significantly higher in 1×OAA than the SAA group and the control(p＜0.05);There were not significantly different in the rate of 4-cell embryo,morula,the blastocyst and hatched blastocyst among SAA group,OAA group and the control(p＞0.05),and the numbers of OAA was higher than the others.(2) The different levels(1,0.5,0.25,0) of OAA were examed,the results:the rates of 2-cell embryo was 89.51%,78.36%,74.62%and 74.68%,respectively.1×OAA group was significantly higher than the 0.25×OAA and 0×OAA group(p＜0.05);the rate of 4-cell embryo was 66.28%,71.27%,71.61%and 52.50%,respectively.0.5 and 0.25 groups were significantly higher than OAA free group(p＜0.05);the rate of morula was 44.59%, 58.77%,50.96%and 41.78%respectively,0.5 group was significantly higher than OAA free group(p＜0.05);the rate of blastocyst was 29.09%,41.71%,33.26%and 22.18%, respectively,0.5 group was significantly higher than the 0 and 1 groups(p＜0.05);there were no significant difference among other rates of different embryo stages in four groups(p＞0.05),0.5 group yielded the best consequence in different embryo stages except for the 2-cell embryo stage.(3) We cultured the embryo step by step according to two above results,such as SAA, 0.5×OAA and 1×OAA after 24 h in 1×OAA,the results showed the rate of blastocyst was 13.23%,28.46%and 23.77%,respectively,0.5 group was significantly higher than SAA group(p＜0.05);there were no significant difference among other rates of different embryo stages in three groups(p＞0.05),and the 0.5 group was the highest one among three groups.(4) We added 25μL HTF solution including 1×ONEAA,1×X3,2×OEAA after culturing the HTF solution including ONEAA,X3(25μL/drop) for 24 h,48 h and 72h.We found that the morula rate was 41.99%,50.76%and 61.53%,respectively,the 72h group was significantly higher than the 24h group(p＜0.05);the rate of blastocyst was 21.69%, 35.64%and 45.26%,respectively,72h group was extremely significantly higher than the 24h group(p＜0.01),48h group was significantly higher than the 24h group(p＜0.05);the rate of hatched blastocyst was 4.44%,10.71%and 22.49%,respectively,72h group was significantly higher than the 24h group(p＜0.01),at the same time,significantly higher than the 48h group(p＜0.05);there were no significant difference among other rates of different embryo stages in three groups(p＞0.05),and the rate of 72h was the highest one.The OEAA had blocked the development of pre-morula embryo,and was benefit to the development of post-morula embryo.In conclusion,in the process of 1-cell embryo culture,it should be selected the OAA as a supplement.After cultured in HTF solution to morula for 72h which including 1×glutamine,arginine,taurine and ONEAA and added 1×OEAA,it could get the best culture results.
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