Palma G A, Müller M, Brem G
Bavarian Research Center in Reproductive Biology, BFZF, Badersfeld, Germany.
J Reprod Fertil. 1997 Jul;110(2):347-53. doi: 10.1530/jrf.0.1100347.
This study was carried out to determine the effects of oestrous cow serum containing insulin-like growth factor I (IGF-I) and supplementation with recombinant IGF-I on subsequent development of bovine embryos produced in vitro. When culture medium was supplemented with oestrous cow serum containing 34.8 ng endogenous IGF-I ml-1, more embryos (P < 0.01) developed to blastocysts by day 9 and more blastocysts hatched on day 13 after insemination (P < 0.01) than in the control group. The effect of the addition of 10, 50 and 100 ng IGF-I ml-1 to culture media containing oestrous cow serum and granulosa cells was also evaluated. Supplementation with 10 ng IGF-I ml-1 did not improve embryo development at any stage. The addition of 50 and 100 ng IGF-I ml-1 did not affect development during the first three cell divisions. However, on day 7 these groups yielded a higher embryo rate than did the control group. Similarly, the proportion of blastocysts on day 9 was enhanced. The addition of 100 ng IGF-I ml-1 also increased the proportion of blastocysts. These data suggest that IGF-I at high concentrations accelerates the development to the blastocyst stage by shortening the transition from the morula to the blastocyst stage. The addition of 100 ng IGF-I ml-1 increased the proportion of hatched blastocysts on day 13. The addition of oestrous cow serum and IGF-I to TCM 199 free of granulosa cells did not increase the proportion of embryos on day 7. However, the progress to blastocysts and hatched blastocysts on days 9 and 13 was significantly lower (P < 0.05). The addition of IGF-I to culture medium without oestrous cow serum but with granulosa cells resulted in significantly lower embryo development than in the control group or in the group supplemented with oestrous cow serum and IGF-I (P < 0.01). The results support the hypothesis that culture media containing high concentrations of IGF-I combined with oestrous cow serum and granulosa cells can improve the development of embryos produced in vitro.
本研究旨在确定含胰岛素样生长因子I(IGF-I)的发情母牛血清以及补充重组IGF-I对体外生产的牛胚胎后续发育的影响。当培养基中添加含34.8 ng内源性IGF-I/ml的发情母牛血清时,与对照组相比,到第9天时有更多胚胎发育至囊胚阶段(P<0.01),且在授精后第13天有更多囊胚孵化(P<0.01)。还评估了向含发情母牛血清和颗粒细胞的培养基中添加10、50和100 ng IGF-I/ml的效果。添加10 ng IGF-I/ml在任何阶段均未改善胚胎发育。添加50和100 ng IGF-I/ml在前三次细胞分裂期间未影响发育。然而,在第7天,这些组的胚胎率高于对照组。同样,第9天囊胚的比例有所提高。添加100 ng IGF-I/ml也增加了囊胚的比例。这些数据表明,高浓度的IGF-I通过缩短从桑椹胚到囊胚阶段的过渡加速了发育至囊胚阶段。添加100 ng IGF-I/ml增加了第13天孵化囊胚的比例。向不含颗粒细胞的TCM 199中添加发情母牛血清和IGF-I在第7天并未增加胚胎比例。然而,在第9天和第13天发育至囊胚和孵化囊胚的进程显著较低(P<0.05)。向不含发情母牛血清但含颗粒细胞的培养基中添加IGF-I导致胚胎发育明显低于对照组或添加发情母牛血清和IGF-I的组(P<0.01)。结果支持这样的假设,即含有高浓度IGF-I并结合发情母牛血清和颗粒细胞的培养基可改善体外生产胚胎的发育。