| This experiment were mainly conducted to bovine oocytes in vitro maturation, in vitro fertilization as well as the earlier embryos cultured in different medium in vitro condition,To find out the influence factor of the embryos producted in vitro.Used aspiration and surface cutting respectively to compare the effects of recovering method on the quality and the quantity of oocytes from ovaries.The recovery rate of aspiration was 86.19% which was higher than surface cutting(78.46%).The percentages of oocytes of A grade for aspiration was significantly higher than surface cutting (p＜0.01).but aspiration got much more oocytes of C grade (p＜0.05).The ovaries which collected from slaughter houses had been divided to two kinds, according to with or without corpus luteum. 20 ovaries had Chosen to observe the effects of corpus luteum on the quality and the quantity of oocytes. The average total number of oocytes for the ovaries with corpus luteum was 11,were remarkably fewer than the ovaries without corpus luteum(20) (p＜0.01).In the same culture condition, The maturation rate of oocytes from the ovaries whitout corpus luteum is 84.33%, significantly higher than the oocytes from the ovaries with corpus luteum on(74.32%) (p＜0.05).The available rates of oocytes had no significant difference in the two kinds ovaries.Added 10%NCS(the comparison group),10%NCS+10% BFF(the 1st group), 10%NCS+20% BFF(the 2nd group), 10%NCS+30% BFF(the 3rd group), 10%NCS+40% BFF(the 4th group), 10% BFF(the 5th group), 20% BFF(the 6th group), 30% BFF(the 7th group) in the TCM-199nutrient fluid respectively.In the 2nd group, the maturation rate(84.93%) were significantly higher (p＜0.01) than the 1st group which added 10%NCS only. The maturation rate drop tendency along with the BFF (+10%NCS) dosage increase when the dosage exceed;when added BFF alone,the best dosage is 20%BFF.The oocytes collected from ovaries 2-6mm follicle were surface cut,dissection was used to collect in-depth oocyte in the ovary(IO).compared the two types of oocytes in IVM,IVC. The maturation rate was 58.20%,and55.84%,respectively. The cleavage rate of oocytes from othe ovary surface 2-6mm follicle was 54.76%,The cleavage rate of 10 was 48.53% The were no significantly difference between two kind of oocytes in the maturation rate and the cleavage rate.Used Cumulus Cells and Bovine Oviductal Epithelial cells(BOEC) co-culture fertilized egg.There were 5 groups in experiment.The 1st group(growth culture medium were supplemented with Cumulus Cells and Bovine Oviductal Epithelial cells(BOEC) in IVM,IVC); The 2nd group(Supplemented with Cumulus Cells in IVM and BOEC in IVC); The 3rd group(added BOEC only in IVM,IVC; The 4th group,added Cumulus Cells only in IVM,IVC). The comparison group(hadn't added Cumulus Cells or BOEC). The 2nd group had got a better result on maturation rate, cleavage rate and developmental rate are 85.90%,65.38%,42.31%, significantly higher than the comparison group (52.38%,34.92%,19.05%) (p＜0.01).and comparatively higher than the 3rd group (p＜0.05).The cleavage rate and blastocyst rate was also higher than the 4th group (p＜0.05). The cleavage rate and blastocyst of third group were hgher than the comparison group.Results demonstrated that surface cutting were ideal method for bovine oocytes collected in IVMFC. The oocytes from the ovaries without corpus luteum got a higher maturing rate in vitro maturation.IO maturation rate and cleavage rate in vitro same with the oocytes from the surface. Added 10%NCS+20%BFF in the medium can improved maturation rate significantly. Cumulus Cells have had promotion in bovine oocytes maturation in vitro, BOEC was a ideal co-culture system that can enhance cleavage rate of fertilized ovum, Supplemented with Cumulus Cells in IVM and BOEC in IVC was the ideal medium for bovine oocytes in IVMFC. |
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