Moore K, Bondioli K R
Granada Biosciences, Inc., College Station, Texas 77480.
Biol Reprod. 1993 Apr;48(4):833-40. doi: 10.1095/biolreprod48.4.833.
One-cell cattle embryos were prepared by in vitro oocyte maturation and fertilization (IVM/IVF) and cultured with or without oviductal cells. Embryos were evaluated after 7 days in culture to determine the percentage developing from the 1-cell stage to the morula or blastocyst stage. The combination of glycine (2 mM) and alanine (1 mM) with oviductal cells (experiment 1) improved embryo development over that in control culture (29 vs. 13%; p < 0.05). An optimum response was obtained with 10 mM glycine and 1 mM alanine in coculture (experiments 2 and 3). In experiment 4, the effects of glycine (0 or 10 mM), alanine (0 or 1 mM), and the presence or absence of oviductal cells were tested. In the absence of oviductal cells, the addition of glycine, alanine, or glycine and alanine combined improved embryonic development over that in control medium (45, 33, 42 vs. 24%, p < 0.001, p < 0.05, p < 0.001, respectively). However, the effect of the combination of glycine and alanine was not different from that of glycine alone when oviductal cells were absent (42 vs. 45%, p > 0.10). In the presence of oviductal cells, glycine or the combination of glycine and alanine improved embryonic development over that in the control medium with cells (47, 55 vs 37%, p < 0.01, respectively). However, supplementation with alanine alone gave no improvement over controls when oviductal cells were present (40 vs. 37%, p > 0.10). These results indicate that glycine and alanine, when used independently, directly affect cattle embryo development, but in combination affect embryo development indirectly, possibly by altering oviductal cell function.(ABSTRACT TRUNCATED AT 250 WORDS)
通过体外卵母细胞成熟和受精(IVM/IVF)制备单细胞牛胚胎,并在有或无输卵管细胞的情况下进行培养。培养7天后对胚胎进行评估,以确定从单细胞阶段发育到桑葚胚或囊胚阶段的百分比。甘氨酸(2 mM)和丙氨酸(1 mM)与输卵管细胞联合使用(实验1)比对照培养更能促进胚胎发育(29%对13%;p<0.05)。共培养中10 mM甘氨酸和1 mM丙氨酸可获得最佳反应(实验2和3)。在实验4中,测试了甘氨酸(0或10 mM)、丙氨酸(0或1 mM)以及有无输卵管细胞的影响。在无输卵管细胞时,添加甘氨酸、丙氨酸或甘氨酸与丙氨酸联合使用比对照培养基更能促进胚胎发育(分别为45%、33%、42%对24%,p<0.001、p<0.05、p<0.001)。然而,在无输卵管细胞时,甘氨酸与丙氨酸联合使用的效果与单独使用甘氨酸无差异(42%对45%,p>0.10)。在有输卵管细胞时,甘氨酸或甘氨酸与丙氨酸联合使用比有细胞的对照培养基更能促进胚胎发育(分别为47%、55%对37%,p<0.01)。然而,在有输卵管细胞时,单独添加丙氨酸与对照相比无改善(40%对37%,p>0.10)。这些结果表明,甘氨酸和丙氨酸单独使用时直接影响牛胚胎发育,但联合使用时可能通过改变输卵管细胞功能间接影响胚胎发育。(摘要截断于250字)