不同添加物对牛卵母细胞体外成熟、核移植胚胎发育及牛卵母细胞冷冻后发育的影响
摘要
一、在卵母细胞成熟液中分别添加EGF、OCS、FBS、BFF和VE;并通过胞质内注射法构建牛体细胞核移植胚胎,研究其对卵母细胞核成熟和胞质成熟的影响;结果如下:
1.在成熟液中添加EGF显著的提高了卵母细胞的成熟率(P<0.05)、极显著提高了核移植胚的卵裂率(P<0.01)和囊胚率(P<0.01)。
2.在成熟液中分别添加OCS(0d)和OCS(7d),OCS(0d)组卵母细胞的成熟率极显著高于OCS(7d)组(P<0.01);OCS(0d)组的卵裂率高于添加OCS(7d)组,但二者之间差异不显著(P>0.05);OCS(0d)组的囊胚率极显著高于OCS(7d)组(P<0.01)。
3.在成熟液中分别添加OCS (0d)和FBS,OCS (0d)组卵母细胞的成熟率、核移植胚的卵裂率及囊胚率均极显著高于FBS组(P<0.01)。
4.在成熟液中分别添加5%BFF+5%FBS和10%BFF,5%BFF+5%FBS组卵母细胞的成熟率显著高于FBS组和10%BFF组(P<0.05),10%组在核移植胚胎的构建过程中容易发生粘连现象,故胚胎未构建,5%+5%FBS组的卵裂率达到了84.61%,囊胚率达到了14.29%。
5.成熟液中添加VE组卵母细胞的成熟率高于对照组,但差异不显著(P>0.05);核移植胚胎的卵裂率和囊胚率均高于对照组,且差异极显著(P<0.01)。
二、在成熟液中分别添加EGF、OCS和VE培养的卵母细胞和对照组卵母细胞分别冷冻保存,卵母细胞解冻后,应用形态正常率和体外受精胚的发育评价培养液各种添加物对卵母细胞的冷冻的影响。结果如下:
1.对照组和添加EGF组的卵母细胞形态正常率分别为71.9%和76.2%,差异不显著(P>0.05),两组的卵裂率分别为63.4%和79.7%,差异显著(p﹤0.05),两组的囊胚率分别为15.1%和30.3%,差异显著(p﹤0.05)。
2.对照组和添加OCS组的卵母细胞形态正常率分别为70.5%和75.7%,差异不显著(P>0.05),两组的卵裂率分别为65.2%和81.4%,差异显著(p﹤0.05),两组的囊胚率分别为13.3%和26.7%,差异显著(p﹤0.05)。
3.对照组和添加VE组的卵母细胞形态正常率分别为73.9%和79.2%,差异不显著(P>0.05),两组的卵裂率分别为60.8%和68.6%,差异不显著(P>0.05),两组的囊胚率分别为14.1%和18.4%,差异不显著(P>0.05)。
Effect of addition of EGF、OCS、FBS、BFF and VE in culture medium on the in vitro maturation of bovine oocytes,then forming bovine SNT embryos by the microinjection of cytoplasm ,study the influce of nuclear and cytoplasm maturation,the results showed:
1. Adding EGF to OM media raising the rate of oocytes’maturation(P<0.05) and NT embryos’cleavage(P<0.01) and blastodysts(P<0.01).
2. Adiding OCS(0d) or OCS(7d) to OM,the rate of maturation in group of OCS(0d) were higher significantly than that of OCS(7d)(P<0.01);the rate of cleavage in group of OCS(0d) is higher than than of OCS(7d) (P>0.05); the rate of blastocysts in group of OCS(0d) were higher significantly than that in group of OCS(7d)(P<0.01).
3. Adding OCS(0d) or FBS to OM,the rates of maturation,cleavage and blastocysts in group of OCS(0d) were all higher significantly than that of FBS(P<0.01).
4. Adding 5%BFF+5%FBS or 10%BFF to OM, the rate of maturation in group of 5%BFF+5%FBS were higher significantly than that of 10%BFF(P<0.05);the rate of cleavage in group of 5%BFF+5%FBS is 84.61%,the rate of blastocysts is 14.29%.
5. Adding VE or not to OM,the rates of maturation is higher than that of control(P>0.05),cleavage and blastocysts in group of VE were all higher than that of control(P<0.01).
Cryopreservation oocytes that treatment in OM adding EGF,OCS and VE or not separately,then thawing oocytes,evaluation the influence of freezing oocytes that different additive in OM using the rate of oocytes’eumorphism and development capacity IVF embryo. the results showed:
1. The rate of oocytes’eumorphism in control group and that adding EGF were 71.9% and 76.2% separately(P>0.05),the rate of cleavage in two groups were 63.4% and 79.7% separately(p﹤0.05),the rate of blastocysts in two groups were 15.1% and 30.3% separately.
2. The rate of oocytes’eumorphism in control group and that adding OCS were 70.5% and 75.7% separately(P>0.05),the rate of cleavage in two groups were 65.2% and 81.4% separately(p﹤0.05),the rate of blastocysts in two groups were 13.3% and 26.7% separately(p﹤0.05).
3. The rate of oocytes’eumorphism in control group and that adding VE were 73.9% and 79.2% separately(P>0.05),the rate of cleavage in two groups were 60.8% and 68.6% separately(p>0.05),the rate of blastocysts in two groups were 14.1% and 18% separately(p>0.05).
1.在成熟液中添加EGF显著的提高了卵母细胞的成熟率(P<0.05)、极显著提高了核移植胚的卵裂率(P<0.01)和囊胚率(P<0.01)。
2.在成熟液中分别添加OCS(0d)和OCS(7d),OCS(0d)组卵母细胞的成熟率极显著高于OCS(7d)组(P<0.01);OCS(0d)组的卵裂率高于添加OCS(7d)组,但二者之间差异不显著(P>0.05);OCS(0d)组的囊胚率极显著高于OCS(7d)组(P<0.01)。
3.在成熟液中分别添加OCS (0d)和FBS,OCS (0d)组卵母细胞的成熟率、核移植胚的卵裂率及囊胚率均极显著高于FBS组(P<0.01)。
4.在成熟液中分别添加5%BFF+5%FBS和10%BFF,5%BFF+5%FBS组卵母细胞的成熟率显著高于FBS组和10%BFF组(P<0.05),10%组在核移植胚胎的构建过程中容易发生粘连现象,故胚胎未构建,5%+5%FBS组的卵裂率达到了84.61%,囊胚率达到了14.29%。
5.成熟液中添加VE组卵母细胞的成熟率高于对照组,但差异不显著(P>0.05);核移植胚胎的卵裂率和囊胚率均高于对照组,且差异极显著(P<0.01)。
二、在成熟液中分别添加EGF、OCS和VE培养的卵母细胞和对照组卵母细胞分别冷冻保存,卵母细胞解冻后,应用形态正常率和体外受精胚的发育评价培养液各种添加物对卵母细胞的冷冻的影响。结果如下:
1.对照组和添加EGF组的卵母细胞形态正常率分别为71.9%和76.2%,差异不显著(P>0.05),两组的卵裂率分别为63.4%和79.7%,差异显著(p﹤0.05),两组的囊胚率分别为15.1%和30.3%,差异显著(p﹤0.05)。
2.对照组和添加OCS组的卵母细胞形态正常率分别为70.5%和75.7%,差异不显著(P>0.05),两组的卵裂率分别为65.2%和81.4%,差异显著(p﹤0.05),两组的囊胚率分别为13.3%和26.7%,差异显著(p﹤0.05)。
3.对照组和添加VE组的卵母细胞形态正常率分别为73.9%和79.2%,差异不显著(P>0.05),两组的卵裂率分别为60.8%和68.6%,差异不显著(P>0.05),两组的囊胚率分别为14.1%和18.4%,差异不显著(P>0.05)。
Effect of addition of EGF、OCS、FBS、BFF and VE in culture medium on the in vitro maturation of bovine oocytes,then forming bovine SNT embryos by the microinjection of cytoplasm ,study the influce of nuclear and cytoplasm maturation,the results showed:
1. Adding EGF to OM media raising the rate of oocytes’maturation(P<0.05) and NT embryos’cleavage(P<0.01) and blastodysts(P<0.01).
2. Adiding OCS(0d) or OCS(7d) to OM,the rate of maturation in group of OCS(0d) were higher significantly than that of OCS(7d)(P<0.01);the rate of cleavage in group of OCS(0d) is higher than than of OCS(7d) (P>0.05); the rate of blastocysts in group of OCS(0d) were higher significantly than that in group of OCS(7d)(P<0.01).
3. Adding OCS(0d) or FBS to OM,the rates of maturation,cleavage and blastocysts in group of OCS(0d) were all higher significantly than that of FBS(P<0.01).
4. Adding 5%BFF+5%FBS or 10%BFF to OM, the rate of maturation in group of 5%BFF+5%FBS were higher significantly than that of 10%BFF(P<0.05);the rate of cleavage in group of 5%BFF+5%FBS is 84.61%,the rate of blastocysts is 14.29%.
5. Adding VE or not to OM,the rates of maturation is higher than that of control(P>0.05),cleavage and blastocysts in group of VE were all higher than that of control(P<0.01).
Cryopreservation oocytes that treatment in OM adding EGF,OCS and VE or not separately,then thawing oocytes,evaluation the influence of freezing oocytes that different additive in OM using the rate of oocytes’eumorphism and development capacity IVF embryo. the results showed:
1. The rate of oocytes’eumorphism in control group and that adding EGF were 71.9% and 76.2% separately(P>0.05),the rate of cleavage in two groups were 63.4% and 79.7% separately(p﹤0.05),the rate of blastocysts in two groups were 15.1% and 30.3% separately.
2. The rate of oocytes’eumorphism in control group and that adding OCS were 70.5% and 75.7% separately(P>0.05),the rate of cleavage in two groups were 65.2% and 81.4% separately(p﹤0.05),the rate of blastocysts in two groups were 13.3% and 26.7% separately(p﹤0.05).
3. The rate of oocytes’eumorphism in control group and that adding VE were 73.9% and 79.2% separately(P>0.05),the rate of cleavage in two groups were 60.8% and 68.6% separately(p>0.05),the rate of blastocysts in two groups were 14.1% and 18% separately(p>0.05).
引文
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