绵羊体外受精胚胎培养系统优化与囊胚差异染色

【目的】评定SOF和CR1培养系统对绵羊体外受精胚胎的发育的影响,筛选适合绵羊体外受精胚胎发育的培养体系,为相关试验研究提供一定的理论依据和研究基础。【方法】通过分析研究SOF和CR1两种培养系统在添加不同蛋白和血清成分时对绵羊体外受精胚胎发育的影响,并采用一种简化的差异染色方法,对SOFaaBSA和CR1aaBSA-FBS组培养液培养的绵羊扩张囊胚和孵化囊胚的总细胞数,滋养层细胞与内细胞团细胞数的组成和比例进行深入比较。【结果】SOFaaBSA和CR1aaBSA-FBS组的卵裂率、囊胚率和囊胚孵化率均高于其它实验组,但这两组之间差异不显著(P0.05)。同时,SOFaaBSA组的扩张囊胚和孵化囊胚的总细胞数和滋养层细胞(TE)数均显著高于CR1aaBSA-FBS组(P0.05),但内细胞团(ICM)数和ICM与TE细胞的比例在两组培养系统中均无显著差异(P0.05)。【结论】CR1aaBSA-FBS培养液能够像SOFaaBSA培养液一样支持绵羊体外受精胚胎的发育,CR1培养系统可以用于绵羊体外受精胚胎的生产,但SOFaaBSA培养液相比较CR1aa BSA-FBS培养液而言更加适合绵羊体外受精胚胎的发育。

Optimization of Culture System and Bastaca Differential Cell Staining of in Vitro Fertilization Embryos of Sheep

【Objective and Mehtod】The aim of the present study was to investigate the respective ability of synthetic oviductal fluid(SOF) and Charles Rosenkrans medium(CR1) media supplemented with FBS or BSA to support in vitro development of sheep IVP embryos and qualities of embryos by differential cell staining.Sheep presumptive IVF zygotes were cultured in the following six media:(1) SOF supplemented with amino acids(SOFaa) and 8 mg/ml bovine serum albumin(BSA) for nine days(SOFaaBSA);(2) SOFaa supplemented with 10% fetal bovine serum(FBS) for nine days(SOFaaFBS);(3) SOFaaBSA for first three days and then SOFaaFBS for later six days(SOFaaBSA-FBS);(4) CR1 supplemented with amino acids(CR1aa) and 8 mg/ml BSA for nine days(CR1aaBSA);(5) CR1aa supplemented with 10% FBS for nine days(CR1aaFBS);(6) CR1aaBSA for first three days and then CR1aaFBS for later six days(CR1aaBSA-FBS).Then,the qualities of embryos were evaluated by differential cell staining of ICM to TE cell in expanded or hatched blastocysts.【Result】The rates of blastaca and hatched blastocyst in group 1,group 3 and group 6 were not different(P> 0.05),but were greater than that in other three groups(P < 0.05).In SOF and CR1 cultural system,SOFaaBSA and CR1aaBSA-FBS