牛体外受精卵发育培养条件的研究

将采自屠宰母牛卵巢的未成熟卵子在含有促进性腺激素和胎牛血清的TCM199培养基内培养成熟后,使用以钙离子载体Ionophore A23187诱导获能的牛精子进行授精处理.在受精卵发育用培养基中添加了谷氨酰胺,并以此与无添对照进行了对比,探索了培养基内谷氨酰胺的存在与否对胚胎发育的影响.另外,在发育用培养基内还分别添加了胎牛血清(FCS),阉牛血清(NSS)以及发情母牛血清(OCS)以观察其培养效果,结果表明,谷氨酰胺对胚胎的发育有极大的促进作用,添加谷氨酰胺的处理组与对照组中发育为桑椹胚、囊胚的胚胎比例分别为23.4%和6.6%,具有极显著的整异.而在含有FCS、NSS和OCS的TCM199培养基内培养的卵子中分别有81.4%,70.7%和82.9%发育为二细胞期胚胎,将胚胎继续培养至授精后第8天时,三个处理组中发育为桑椹胚、囊胚的胚胎比例分别为30.0%,40.2%和47.1%,以OCS的培养效果为最佳.本研究的结果证明,谷氨酰胺和发情母牛血清对牛体外受精卵的体外发育具有良好的促进作用.

The Study on the Culture Conditions of in vitro Fertilized Bovine Embryos

The oocytes recovered from slaughtered cow were cultured in M199 supplemented with fetal calf serum and gonadotropin,then the matured oocytes were inseminated with bull semen treated with Ionophore A23187.The effect of glutamin and different sera supplemented in the medium on the development of IVF embryos were examined.The results showed that glutamine exerted a beneficial effect on embryo development.The culturing of IVF embryos in M199 medium supplemented with glutamine allowed 23. 4% of embryos developed to morulae and blastocysts,but the development rate in control group was only 6. 6% (P<0.01).The embryos cultured in the M199 supplemented with fetal calf serum (FCS),normal steer serum (NSS)and oestrus cow serum (OCS)could develop to morulae and blastocysts with the percentage of 30.0%,40.2% and 47.1%,respectively.OCS has the best effect on embryo development.The results of present study proved that glutamine and cestrus cow serum could promote the development of the IVF embryos cultured in vitro.