牛体外受精早期胚胎培养体系优化的研究

Optimization Study of the Early Embryo Culture System of Bovine in Vitro Fertilization

研究旨在筛选牛体外受精的最优培养体系,提高体外受精的囊胚孵化率。本研究将培养体系分为四组,CR1aa组,mCR组,CR1aa前+mCR后组,mCR前+CR1aa后,通过统计胚胎发育情况。结果显示CR1aa组卵裂率,八细胞率,囊胚率,孵化率分别为82.29±0.58,48.79±1.12,24.99±0.28,14.42±0.86;mCR组卵裂率,八细胞率,囊胚率,孵化率分别为57.18±1.61,26.48±0.53,22.15±1.20,18.12±0.57;CR1aa前+mCR后组卵裂率,八细胞率,囊胚率,孵化率分别为82.24±1.32,48.60±1.93,34.07±1.04,24.87±0.66;mCR前+CR1aa后组卵裂率,八细胞率,囊胚率,孵化率分别为58.80±2.26,26.77±1.21,24.15±0.73,13.10±1.09;从四组结果中可以看出CR1aa前培养液与mCR前相比可以显著提高牛体外受精的卵裂及八细胞率(P<0.05),但其后期发育不佳。mCR后与CR1aa后相比可以显著提高牛体外受精胚胎的囊胚率和孵化率(P<0.05)。由此得出mCR前+CR1aa后组可以显著的提高体外受精的囊胚和孵化率。为了进一步验证该组合的稳定性,利用mCR前+CR1aa后体系分别在不同季节进行验证,结果显示除夏季相对不佳外,其他季节均能维持较高的卵裂囊胚孵化率。进而认为CR1aa前+mCR后体系最有利于牛体外受精胚胎的生产且孵化率较高,有利于后期胚胎移植的进行。

In order to further find a optimal culture system of IVF and improve the blastocyst incubation rate in IVF.In this study,CR1aa group,mCR group,pre-CR1aa+ post-mCR group and pre-mCR + post-mCR group were programmed.The results showed that the cleavage rate,eight cell rate,blastocystrate and hatching rate of CR1aa group were 82.29±0.58,48.79±1.12,24.99±0.28,14.42±0.86;mCR group group were 57.18±1.61,26.48±0.53,22.15±1.20,18.12±0.57;and preCR1aa+ post-mCR group were 82.24±1.32,48.60±1.93,34.07±1.04,24.87±0.66;pre-mCR +post-mCR were 58.80±2.26,26.77±1.21,24.15±0.73,13.10±1.09;It can be seen from the results of four groups that compared with mCR,CR1aa anterior culture solution can significantly improve the cleavage and eight cell rate(P<0.05)of bovine in vitro fertilization,but its late development is poor.The blastocyst rate and hatching rate(P<0.05)of bovine in vitro fertilization embryos could be significantly increased post-mCR.It is concluded that the pre-CR1aa+post-mCR can significantly improve the cleavage blastocyst and hatching rate of in vitro fertilization.In order to further verify the stability of the combination,the results of the pre-CR1aa+ post-mCR applied in different seasons showed that the hatching rate of the cleavage blastocyst could be maintained in other seasons except for the relatively poor summer.Our study found that the pre-CR1aa+post-mCR system provided a stable culture system for the production of bovine IVF embryos and a high incubation rate,which was conducive to the subsequent embryo transfer.