| The aim of this study was to establish an steady and efficient ICSI system for sheep oocytes maturation in vitro in our lab. Experiments were carried out to analyze the diffetent factors which been reported influence the results of ICSI and to optimize the technology of ICSI in sheep and establish a preliminary IVF system. Results:1.The sperm were treatment by DTT and untreated.after ICSI, The Cleavage rate (79.6%,81.6%;P>0.05)were not significantly different between the two group;whereas,The blastocys rate(31.2%,20.9%;P<0.05)was higher for treatment than untreated. 2.ICSI with live and dead status of the sperm, There was no significant difference of Cleavage rate((80.0%,83.1%)and blastocys rate(27.6%,20.2%) between the two group( P>0.05).3. After ICSI ,the oocytes were activated by different method:(1) Sham-injected, no activated. (2) Sperm-Injected, no activated. (3) Sham-injected, oocytes were activated with Ionomysin+6-DMAP.(4)Sperm-injected, oocytes were activated with Ethanol+6-DMAP.(5) Sperm-injected, oocytes were activated with Ionomysin+6-DMAP. Results : In Sham-injected and Sperm-injected oocytes,but no activated, both the cleavage rate(7.5%,5.3%)and blastocyst rate(0.94%,1.8%)were very low. The cleavage rate(65.7%,71.6%,75.6%;P>0.05)of 3,4,5 groud was no significant difference,but blastocyst rate(5.7%,15.6%,26.0%;P<0.01) of the third groud was lower than others. There was no significant difference of cleavage rate(71.6%,75.6%;P>0.05)of the sperm-injected groud treated with Ethanol+6-DMAP and Ionomysin+6-DMAP,while blastocyst rate (15.6%,26.0%;P<0.05)was lower than latter. 4. This study was to evaluate the Effects of first polar body position on fertilization of sheep oocytes by intracytoplasmic sperm injection .There was no significant difference of cleavage rate (82.4%%,85.8%;P>0.05)and blastocyst rate (28.8%,26.9%;P>0.05 ) to first polar body positioned at 12 o'clock or 6 o'clock. 5. The different concentration(10%,8%,5%) PVP were added to manipulation medium ,The results showed that cleavage rate (80.2%,83.6%,72.0%; P<0.05)and blastocyst rate (27.8%,30.5%,16.0%;P<0.05) was higher for 10% and 8%PVP than 5%PVP . (6) Added VEGF in culture medium , There was no significant difference of cleavage rate (78.7%,83.9%;P>0.05)and blastocyst rate (27.6%,23.6%;P>0.05 ) in the same condition with or without VEGF. (7) There was no significant difference of cleavage rate (77.5%,74.7%;P>0.05)in the same condition with or without CB,but blastocyst rate (30.5%,16.0%;P<0.05) was higher for with than without CB. (8) Injection eggs in room temperature and 37.0℃, respectively. the cleavage rate(90.6%,80.1%;P<0.05)was significantly higher for room temperature than that of latter, but the blastocyst rate(31.3%,28.2%;P>0.05) was not significantly different between the two group. (9) Oocytes were randomly injection for 0-1h,1-2h and 3-4h , cleavage rate were 46.07%,43.33%,30.56% (P>0.05),respectively.The blastocyst rate was higher for oocytes injection for 0~1 h than 1~2 h,3~4 h(33.6%,18.3%,13.0%;P<0.05-0.01), There was no significant difference of blastocyst rate between oocytes injection for 1-2h and 3-4h (P>0.05)...
|
PDF Full Text Request