培养液中血清或其替代物对牛体外受精卵体外发育的影响

探讨在牛体外受精卵体外培养液中添加血清替代品对体外受精卵发育的影响,确定血清在SOF培养液中合适的添加浓度及添加时期,以筛选优良的牛体外受精卵体外培养液.在SOF液中分别添加10%血清(NBS)、8 mg/ml牛血清白蛋白(BSA)和1 mg/ml聚乙烯醇(PVA)进行牛体外受精卵的全程培养,添加NBS组的卵裂率(66.3%)显著高于添加BSA组(51.0%)(P＜0.05),极显著高于添加PVA组(47.6%)(P＜0.01);添加NBS组的囊胚发育率(30.9%)显著高于添加PVA组(20.7%)(P＜0.05),极显著高于添加BSA组(13.6%)(P＜0.01).在培养液中添加不同浓度(10%、5%)的NBS及无血清培养液全程培养时,3组之间卵裂率(65.6%、60.0%、58.0%)差异不显著(P＞0.05);10%NBS组囊胚发育率(30.0%)极显著高于5%NBS组(17.6%)和无血清组(6.0%)(P＜0.01).5%NBS组的囊胚发育率极显著地高于无血清组(P＜0.01).在受精卵培养的最初48 h用5%NBS,之后用10%NBS培养液时,卵裂率(58.6%)和囊胚发育率(29.3%)与SOF+10%NBS全程培养组(66.3%、30.9%)差异都不显著(P＞0.05).试验结果表明,SOF培养液中添加PVA、BSA替代血清时卵裂率和囊胚发育率都明显下降,可能SOF培养液中添加血清比添加PVA或BSA更有利于牛体外受精卵的体外发育.体外培养时在SOF培养液中添加一定浓度的血清以及改变血清的添加方式,可提高牛体外受精卵体外发育效果.

Influences of Serum and Serum Substitutes in the Medium on Bovine IVF Zygotes Development in Vitro

This experiment was designed to determine the effects of serum substitutes addition in bovine IVF zygotes culture medium,and determine the appropriate concentration and the prime time to add serum in SOF medium,in order to get a fine culture medium for bovine IVF zygotes.10%NBS,8 mg/ml BSA and 1 mg/ml PVA were added in SOF medium respectively to culture bovine IVF zygotes.The cleavage rate(66.3%) of the group containing NBS was significantly higher than the groups containing BSA and PVA(51.0%,47.6%)(P<0.05,P<0.01).The bastocyst rate(30.9%) of group containing NBS was significantly higher than the bastocyst rate of groups containing PVA and BSA(20.7%,13.6%)(P<0.05,P<0.01).10% and 5% NBS was added in the culture medium respectively and free NBS for full process culture.There were no significant differences among these three groups on cleavage rate(65.6%,60.0%,58.0%).The blastocyst rate(30.0%) of the group with 10%NBS was significantly higher than groups with 5%NBS(17.6%) and free NBS(6.0%)(P<0.01),and the blastocyst development rate of 5%NBS group was significantly higher than the group free from NBS(P<0.01).When the zygotes were cultured in medium with 5%NBS for the first 48 h,then cultured in the medium with 10%NBS,the cleavage rate(58.6%) and blatocyst development rate(29.3%) were all higher than SOF 10%NBS(66.3%,30.9%) full process cultivation and there was no different between the two groups(P>0.05).The results showed that,when PVA and BSA were added in SOF medium instead of serum,the cleavage rate and blastocyst developing rate descended obviously,it illuminated that may be the addition of serum in SOF medium was better than PVA and BSA for the in vitro development of bovine IVF zygotes.Serum of certain concentration added in SOF medium and change the addition mode can promote the development of bovine IVF zygotes when cultured in vitro.