Thibodeaux J K, Myers M W, Prough S G, White K L
In Vitro Fertilization Laboratory, Tulsa Center for Fertility & Women's Health, Tulsa, OK 74104, USA.
Theriogenology. 1995 Aug;44(3):423-32. doi: 10.1016/0093-691x(95)00196-f.
The present experiments were conducted to determine if supplementation of the culture medium with a serum extender containing growth factors would increase development of bovine embryos into morulae or blastocysts, following in vitro maturation (IVM) and in vitro fertilization (IVF). In Experiment 1, bovine zygotes were cultured in CR1 medium supplemented with 0, 0.01, 0.1, 1 or 10% serum extender. In Experiment 2, bovine zygotes were cultured in the presence of cumulus cells in CR1 medium supplemented with 0, 0.01, 0.1, 1 or 10% serum extender. In Experiment 3, bovine oocytes were matured in Medium 199 supplemented with 0, 0.01, 0.1, 1 or 10% serum extender. In Experiment 4, oocytes were matured in Medium 199 with 10% fetal bovine serum (FBS) or 5% FBS with serum extender. Following maturation, zygotes were cultured in CR1 medium with 10% FBS or 5 % FBS and serum extender. In all 4 experiments, the embryos were cultured in vitro until Day 7 after IVF, and development to the morula or blastocyst stage was assessed. The findings of the first 2 experiments showed that the serum extender did not directly influence embryo development but did stimulate development when cumulus cells were included in the culture system. The remaining 2 experiments showed that the serum extender did influence development through its interactions with cumulus cells during maturation and/or culture. These findings suggest that although growth factors or other products do not directly stimulate bovine embryo development their effects may be mediated through secondary cell systems.
进行本实验是为了确定在体外成熟（IVM）和体外受精（IVF）后，向培养基中添加含有生长因子的血清扩展剂是否会增加牛胚胎发育为桑葚胚或囊胚的比例。在实验1中，将牛受精卵在添加了0%、0.01%、0.1%、1%或10%血清扩展剂的CR1培养基中培养。在实验2中，将牛受精卵在含有卵丘细胞的情况下，在添加了0%、0.01%、0.1%、1%或10%血清扩展剂的CR1培养基中培养。在实验3中，将牛卵母细胞在添加了0%、0.01%、0.1%、1%或10%血清扩展剂的199培养基中成熟。在实验4中，卵母细胞在含有10%胎牛血清（FBS）或5% FBS及血清扩展剂的199培养基中成熟。成熟后，将受精卵在含有10% FBS或5% FBS及血清扩展剂的CR1培养基中培养。在所有4个实验中，胚胎在体外培养至IVF后第7天，并评估其发育至桑葚胚或囊胚阶段的情况。前两个实验的结果表明，血清扩展剂不会直接影响胚胎发育，但当培养系统中包含卵丘细胞时会刺激发育。其余两个实验表明，血清扩展剂在成熟和/或培养过程中通过与卵丘细胞的相互作用影响发育。这些发现表明，尽管生长因子或其他产物不会直接刺激牛胚胎发育，但其作用可能通过次级细胞系统介导。
