Abdoon A S, Kandil O M, Otoi T, Suzuki T
Department of Animal Reproduction and Artificial Insemmination [corrected], National Research Centre, Tahrir Street, Dokki, 12622, Giza, Egypt.
Anim Reprod Sci. 2001 Mar 30;65(3-4):215-23. doi: 10.1016/s0378-4320(01)00079-3.
The present study was designed to examine the influence of oocyte quality, culture media and gonadotropins on cleavage rate and development of in vitro fertilized buffalo embryos. Three experiments were conducted. In experiment 1, oocytes were classified by number of cumulus cell layers and morphology of the ooplasm as good, fair or poor. Oocytes were cultured for IVM, IVF and IVC in CR1aa medium. In experiment 2, good quality oocytes were cultured for maturation in: (1) CR1aa; (2) CR2aa; (3) TCM-199; (4) MEM or (5) RPMI-1640, and then fertilized using frozen thawed buffalo spermatozoa in CR1aa. The oocytes were cultured in the same medium used for maturation after fertilization. In experiment 3, oocytes were classified into three groups: group (1) was without gonadotropin and serve as a control; group (2) in which IVM medium was supplemented with 10microg/ml FSH and group (3) in which IVM medium was supplemented with 10IUml(-1) eCG. In all experiments, oocytes were kept at 38.5 degrees C under 5% CO(2) for IVM, IVF, IVC and examined for cleavage and embryo development rates on days 3 and 8, respectively. Good and fair quality oocytes produced a higher cleavage rate (P<0.01) than poor quality oocytes. Morula production rate was also higher (P<0.01) for good as compared with fair quality oocytes. Embryo development with poor quality oocytes was arrested at the two to sixteen cell stage. In experiment 2, the cleavage rate was higher (P<0.05) in CR1aa than CR2aa, and higher (P<0.01) than TCM-199, MEM and RPMI-1640. The numbers of morulae and blastocysts were higher (P<0.01) for oocytes cultured in CR1aa and CR2aa media than TCM-199 or MEM. In experiment 3, the addition of FSH or eCG to the maturation medium increased (P<0.01) cleavage and developmental rates of buffalo embryo compared with control medium. In conclusion, the IVM of good quality buffalo oocytes in CR1aa or CR2aa medium and the addition of FSH or eCG in maturation medium produced higher cleavage and developmental rates of IVF buffalo embryos.
本研究旨在探讨卵母细胞质量、培养基和促性腺激素对体外受精水牛胚胎的卵裂率及发育的影响。进行了三项实验。在实验1中，根据卵丘细胞层数和卵质形态将卵母细胞分为优质、中等或劣质。卵母细胞在CR1aa培养基中进行体外成熟（IVM）、体外受精（IVF）和体外培养（IVC）。在实验2中，将优质卵母细胞分别在以下培养基中进行成熟培养：（1）CR1aa；（2）CR2aa；（3）TCM - 199；（4）MEM或（5）RPMI - 1640，然后在CR1aa中使用冷冻解冻的水牛精子进行受精。受精后，卵母细胞在用于成熟培养的相同培养基中进行培养。在实验3中，将卵母细胞分为三组：第（1）组不添加促性腺激素作为对照；第（2）组的IVM培养基中添加10μg/ml促卵泡素（FSH）；第（3）组的IVM培养基中添加10IU/ml孕马血清促性腺激素（eCG）。在所有实验中，卵母细胞在38.5℃、5%二氧化碳条件下进行IVM、IVF、IVC，并分别在第3天和第8天检查卵裂和胚胎发育率。优质和中等质量的卵母细胞产生的卵裂率高于劣质卵母细胞（P<0.01）。与中等质量卵母细胞相比，优质卵母细胞的桑葚胚产生率也更高（P<0.01）。劣质卵母细胞的胚胎发育停滞在2 - 16细胞阶段。在实验2中，CR1aa中的卵裂率高于CR2aa（P<0.05），且高于TCM - 199、MEM和RPMI - 1640（P<0.01）。在CR1aa和CR2aa培养基中培养的卵母细胞产生的桑葚胚和囊胚数量高于TCM - 199或MEM（P<0.01）。在实验3中，与对照培养基相比，在成熟培养基中添加FSH或eCG可提高水牛胚胎的卵裂率和发育率（P<0.01）。总之，在CR1aa或CR2aa培养基中对优质水牛卵母细胞进行IVM，以及在成熟培养基中添加FSH或eCG可提高体外受精水牛胚胎的卵裂率和发育率。
