Study On The Effect Of Ovum Pick Up And Embryo Prodution In Vitro In Goat

The major factors affected the Laparoscopic Ovum Pick-up (LOPU) and in vitrofertilization (IVF) technique were discussed systematically in this study in order toestablish an acceptable manipulation and production system of LOPU-IVF in our lab. Theobjective of the present study was to improving the in vitro developmental capacity of goatembryo obtained by LOPU and IVF.The oocytes were collected from slaughtered goat ovaries, in this study, three needlegauge (7,9 and 12 G) were used to compare the influence of different needle gauge onoocytes recovery rate. There was difference (P＜0.05) between the three needle gauge onoocytes recovery rate (52.63ï¼…,66.78ï¼…and 62.41 respectively), and there was significantdifference(P＜0.05) between the three needle gauge on good quality oocyte rate(55ï¼…,63.59ï¼…and 68.82ï¼…,respectively). There was no difference(P＞0.05) between 9 and 12 Gneedle on oocytes recovery rate and oocytes useable rate, so 9 and 12 G needle weresuitable for LOPU. With two weeks interval and repeat three times, compare the influencebetween standard surgery and LOPU on the damage of reproductive organ, there was nodifference(P＞0.05) between the two methods on oocytes recoveryrate(76.47ï¼…,74.01ï¼…)and good quality oocyte rate (75.24ï¼…,73.33ï¼…),but standardsurgery made more damage on reproductive organ, what is more, the third standard surgerycouldn't be successfully done because of the adhesion. Goats were divided into threegroups, treated by LOPU, then compare the influence of different interval time on thedamage of reproductive organ, the result was, one week interval group got 12.08visiblefollicles every goat every time, and 8.75 oocytes every goat every time, it had difference(P＜0.05) with two weeks interval group (15.58,11.97) and three weeks intervalgroup(14.92,11.42),but the three groups had no difference on oocytes recovery rate(72.41ï¼…, 76.47ï¼…, 78.29ï¼…)and good quality oocyte rate(74.28ï¼…, 75.24ï¼…, 74.45ï¼…). Oneweek interval group had more damage on ovary, there were many gores on ovary, andserious adhesion; two weeks interval group, ovaries could almost got well, but had a fewgores and little adhesion; three weeks interval group, ovaries could got well, andsometimes had little adhesion.The supplement of 10～60ng/ml EGF in IVM medium groups(90.67％～88.46ï¼…) had higher polar body rate than without EGF group (75.68ï¼…). Thesupplement of 20ng/ml EGF had the highest polar body rate (92.86ï¼…), but there were no difference between the groups which supplement with EGF. The supplement of EGF inIVM medium improved the developmental capacity of embryo after IVF, blastula ratevaries directly as the increase of the concentration of EGF(0～60ng/ml). Oestrus goat serumcan be replaced by oestrus ewe serum as the sperm capacitation factor the cleavage rate is60.42ï¼…, the supplement of 5ï¼…,10ï¼…,20ï¼…oestrus sheep serum in IVF medium, thecleavage rate were 58.33ï¼…, 59.72ï¼…and 62.50ï¼…respectively, they had significantdifference with IVF medium without oestrus sheep serum(0ï¼…, P＜0.01). The supplementof 10μg/ml and 20μg/ml heparinum in IVF medium, the cleavage rate were 87.69ï¼…and85.71ï¼…respectively, the two groups had no difference, but had significant difference withIVF medium without heparinum(59.02ï¼…, P＜0.01). The supplement of 1ï¼…EAA in IVCmedium(29.17ï¼…) had difference with supplement of 2ï¼…EAA(17.10ï¼…).Embryoscultured in 5ï¼…CO₂ in air or 5ï¼…CO₂, 5ï¼…O₂, and 90ï¼…N₂, the two groups had nodifference in morula rate, but the latter had higher blastula rate (30.16ï¼…vs 16.27ï¼…), it saidthe embryo developmental capacity may be improved with lower percent of oxygen in air.The blastula rate of supplement of 5.10.20.40ng/ml EGF in IVC medium had differencewith IVC medium without EGF, but the blastula rate of supplement of 80ng EGF is 5.50ï¼…,it's lower than IVC medium without EGF(15.29ï¼…, P＜0.05 ), and much lower thansupplement of 5.10.20.40ng/ml EGF in IVC medium(28.75ï¼…, 32.76ï¼…, 31.25ï¼…, 26.78ï¼…,P＜0.01). The oocytes collected from slaughtered goat ovaries had no difference withoocytes collected from LOPU in cleavage rate and morula rate, but they had difference inblastula rate(30.16ï¼…vs 14.71ï¼…P＜0.05). 40 early goat embryos of LOPU-IVF weretransfered to the oviducts of 8 recipients, and 1 recipient was pregnanted by B-ultrasonicpregnancy diagnosis two monthes later after ET, the pregnancy rate is 12.5ï¼….