胚胎分割液及分割方法对绵羊体外胚分割效果的影响

The Effects of Different Embryo Bisection Solutions and Bisection Methods on the Result of in Vitro Embryos Bisection of Ovine

主要研究了胚胎分割液、分割方法和胚胎发育时期对绵羊体外生产胚胎的影响。体外采集绵羊卵丘卵母细胞复合体,成熟培养24 h,经过体外受精培养,得到体外胚胎样品。借助显微操作仪,将体外受精的桑葚胚和囊胚分割,体外培养半胚,观察其发育情况。结果发现:在D-PBS+0.2 mol/L蔗糖液与D-PBS+5%PVP液中分割桑葚胚和囊胚,其分割成功率及半胚的囊胚发育率及囊胚细胞数三者均无差异显著性(P>0.05);用显微分割法和徒手分割法分割桑葚胚和囊胚,其分割成功率、半胚的囊胚发育率及囊胚细胞数均无差异显著性(P>0.05);比较桑葚胚和囊胚分割,囊胚的分割成功率显著高于桑葚胚的分割成功率(P<0.05),而半胚的囊胚发育率及半胚细胞数均无差异显著性(P>0.05)。结果表明:在D-PBS液中分别添加0.2 mol/L的蔗糖和5%的PVP都可作为绵羊体外胚的胚胎分割液;显微分割法和徒手分割法均可用于绵羊体外胚胎的分割;囊胚比桑葚胚更适于胚胎分割。

The effects of different embryo bisection solutions and methods and embryo development stages on in vitro embryos bisection of ovine were researched in this experiment. Ovine cumulus-oocyte complex was matured for 24 hours and fertilised for 17 hours in the vitro culture. The samples of embryos were obtained. The morula and blastocysts were splitted by micromaniputation. The bisection embryos were cultured in vitro, and the developmental capacities were observed. The results showed that the bisection efficiency and blastula developmental rate and blastula population of morula and hlastocsyts spirted in D-PBS ＋0.2 mol/L sucrose solution and D-PBS＋5% PVP solution had no significant difference （P 〉 0.05）, and so compared morula with blastocsyts;the bisection efficiency of morula was significantly higher than that of blastocsyts （P 〈0.05） ; however, the blastula developmental rate and population of splitted embryos had no significant difference （P〉 0.05）. Thus,D-PBS＋0.2 mol/L sucrose solution and D-PBS＋5% PVP solution can be used for in vitro embryos bisection solution of ovine; both the micromanipulation and barehanded bisection methods can be used for in vitro embryos bisection of ovine; however,blastocsyt is more suitable than morula for in vitro embryos bisection.