【作者】 韩雪花；

【导师】 罗光彬；

【作者基本信息】 延边大学 ， 动物遗传育种与繁殖， 2002， 硕士

【摘要】 为了探讨外界因素对牛卵母细胞的体外受精及早期胚胎体外培养的影响，研究了受精温度（37℃，38℃，39℃）、卵子基本液（TCM-199，PBS）、激素（LH、E₂）、卵子等级（A级，B级，C级）、受精卵培养方式（共培养，非共培养）以及受精时间（6h，12h，18h）等对牛卵母细胞体外成熟的影响。其结果如下： 1．不同温度（37℃、38℃及39℃）下进行体外受精时，体外受精率分别为78.0％、78.8％、79.9％；囊胚发育率分别为9.0％、9.3％、17.3％；囊胚孵化率分别为8.1％、8.6％、15.2％。经统计分析各组的体外受精率无显著性差异（P＞0.05）；囊胚发育率是39℃组显著高于37℃组和38℃组（P＜0.05）；囊胚孵化率是39℃组极显著高于37℃组和38℃组（P＜0.01）；37℃组和38℃组的囊胚发育率以及孵化率之间无显著性差异（P＞0.05）。 2．用不同基本培养液进行试验时，TCM-199作为基本培养液的牛卵母细胞的体外受精率、囊胚发育率、囊胚孵化率分别为56.3％、15.4％、10.5％；而以PBS为基本培养液的分别为33.6％、13.8％、8.7％。TCM-199基本培养液液组受精率极显著高于PBS基本培养液组（P＜0.01），而囊胚发育率、囊胚孵化率均无显著性差异（P＞0.05）。 3．不同培养液中添加激素的培养液组的受精率、囊胚发育率及囊胚孵化率分别为81.1％、15.3％、9.6％；无激素培养液组的分别为75.8％、12.1％、10.5％。添加激素组和无激素组的受精率、囊胚发育率及囊胚孵化率均无显著性差异（P＞0.05）， 影响牛卵毋纫胞体外受精及胚胎发育的几种因素的研究——中文摘要 4 卵子等级对胚胎发育的影响较大。A级卵母细胞的受精率、囊 胚发育率以及囊胚孵化率分别为 95．2％、25．5％，22．l％；B级分别 为sl．2％、17．1％及13．7％；C级分别为43石％、9．7％及7．3％。A级 和B级的受精率极显著高于C级炉＜0，01)；卵母细胞的囊胚发育率为 A级极显著高于C级o＜O．01X但B级与C级、A级之间均无显著性 差异（P＞0刀5）；牛卵母细胞的囊胚孵化率为A级极显著高于B级和 C级什＜0刀1)，而B级和C级之间差异不显著（P＞0刀5）。 5．共培养受精卵的受精率、囊胚发育率及囊胚孵化率分别为 抡刀％、门5％、9．9％；而普通方法培养的分别为68二％、5，4％、3石％。 共培养受精卵的受精率、囊胚发育率以及囊胚孵化率显著高于普通培 养方法受精卵的受精率、囊胚发育率及囊胚孵化率。 6．不同受精时间下（6h～18h）进行体外受精时6h组的受精率。 囊胚发育率及囊胚孵化率分别为65．1％、9．1％、8．4％；IZ h组分别为 77刀％、17．8％、15石％；18 h组分别为 67．5％、10．9％、9．8％。12 h 组的受精率与囊胚发育率显著高于6h组和18h组（P＜0刀5），而6h 组与18 h组之间均无显著性差异（P＞0刀5）；囊胚孵化率为12 h组显 著高于 6 h（P＜0．05），而 18 h与 12 h、6 h组均无显著差异（P＞0．05）， 但 12 h组的囊胚孵化率高于 18 h组。更多还原

【Abstract】 In order to discuss the influence of the environmental factors for bovine oocytes in vitro fertilization and early embryo cultivation,the bovine oocytes which were cultured maturely in vitro about 24h,which were contained 5% CO2,95% air,and which were cultured in maximum relative humidity,were researched the temperature of fertilization(37,38,39),the time of fertilization (6 h,12 h,18 h),the method of cultivation of zygotes (co-culture,unco-culture),the level of ova (level A,level B,level C),the class of fundamental fluid of ova (TCM,PBS),and the type of hormone.l.The result displayed that various fertilization temperature were no significance effect(P>0.05) to the rate of zygote cleavage,the developmental rate and the hatching rate of blasrula in vitro were submitted elevated tendency accompanied with the temperature of fertilization. The developmental rate and hatching rate of blastulas in 39 C were significance higher than those of blastulas in 38or 37,respectively (p<0.05) .2.In comparison of the two types of the fundamental solution of ova fertilized rate,the bundle which solution TCM was uesed was extremely higher than the bundle which solution PBS was used (p<0.01),but there were no significance difference in the developmental rate and hatchingrate of blastula(P>0.05).3.There was no significance difference between added hormone or not(P>0.05),when researching the effect of hormone to fertilized rate,but there were no significance difference in developmental rate and hatching rate of blastula (p>0.05) .4.In various levels of ova there were extreme difference between level A and level B,level B and level C,respectively (p<0.01),and there were no significance difference among the others (p>0.05),while the hatching rate of blastula,there were extreme difference between level A and level B,level A and level C (p<0.01),while there were no significance difference among the others(P>0.05).5.In various culturing method,the fertilized rate,the developmental rate,and the hatching rate of blastula of co-culruring method were significancely higher than that of unco- culturing method (p<0.05) .6.In various fertilized time,the fertilized rate and the developmental rate of blastula at 12 h,are significance higher than those of blastula at 6 hor 18h,respectively (p<0.05),while there was no significance difference between 12h and 18h (p>0.05). The hatching rate of blastula at 12h were significance higher than that of blastula at 6 h(p<0.05),which there were no significance differences between 12 h and 18 h,6 h and 18h,respectively (p>0.05).更多还原