Laboratoire de Pathologie de la Reproduction et Biotechnologie animale, Ecole Nationale Vétérinaire de Nantes, Nantes, France.
Theriogenology. 2010 Mar 15;73(5):595-604. doi: 10.1016/j.theriogenology.2009.10.015. Epub 2009 Dec 24.
This study examined the influence of the following growth factors and cytokines on early embryonic development: insulin-like growth factors I and II (IGF-I, IGF-II), basic fibroblast growth factor (bFGF), transforming growth factor (TGF-beta), granulocyte-macrophage colony-stimulating factor (GM-CSF), and leukemia inhibitory factor (LIF). Synthetic oviduct fluid (SOF) was used as the culture medium. We studied the development of bovine embryos produced in vitro and cultured until Day 9 after fertilization. TGF-beta1, bFGF, GM-CSF, and LIF used on their own significantly improved the yield of hatched blastocysts. IGF-I, bFGF, TGF-beta1, GM-CSF, and LIF significantly accelerated embryonic development, especially the change from the expanded blastocyst to hatched blastocyst stages. Use of a combination of these growth factors and cytokines (GF-CYK) in SOF medium produced higher percentages of blastocysts and hatched blastocysts than did use of SOF alone (45% and 22% vs. 24% and 12%; P<0.05) on Day 8 after in vitro fertilization and similar results to use of SOF+10% fetal calf serum (38% and 16%, at the same stages, respectively). The averages of total cells, inner cell mass cells, and trophectoderm cells of exclusively in vitro Day-8 blastocysts for pooled GF-CYK treatments were higher than those for SOF and similar to those for fetal calf serum. The presence of these growth factors and cytokines in the embryo culture medium therefore has a combined stimulatory action on embryonic development; in particular through an increase in hatching rate and in the number of cells of both the inner cell mass and trophoblast. These results are the first to demonstrate that use of a combination of recombinant growth factors and cytokine, as IGF-I, IGF-II, bFGF, TGF-beta1, LIF, and GM-CSF, produces similar results to 10% fetal calf serum for the development of in vitro-produced bovine embryos. This entirely synthetic method of embryo culture has undeniable advantages for the biosecurity of embryo transfer.
胰岛素样生长因子 I 和 II（IGF-I、IGF-II）、碱性成纤维细胞生长因子（bFGF）、转化生长因子（TGF-β）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白血病抑制因子（LIF）。使用合成输卵管液（SOF）作为培养基。我们研究了体外生产的牛胚胎的发育，并培养至受精后第 9 天。TGF-β1、bFGF、GM-CSF 和 LIF 单独使用时显著提高了孵化囊胚的产量。IGF-I、bFGF、TGF-β1、GM-CSF 和 LIF 显著加速了胚胎发育，尤其是从扩张囊胚到孵化囊胚阶段的变化。在 SOF 培养基中使用这些生长因子和细胞因子的组合（GF-CYK）比单独使用 SOF 产生更高比例的囊胚和孵化囊胚（受精后第 8 天分别为 45%和 22%对 24%和 12%；P<0.05），并且与使用 SOF+10%胎牛血清的结果相似（分别为 38%和 16%，在相同阶段）。单独在体外培养第 8 天的囊胚中，仅 GF-CYK 处理的囊胚总细胞数、内细胞团细胞数和滋养外胚层细胞数平均值高于 SOF，与胎牛血清相似。因此，这些生长因子和细胞因子在胚胎培养液中的存在对胚胎发育具有协同刺激作用；特别是通过提高孵化率和内细胞团和滋养外胚层细胞的数量。这些结果首次证明，使用 IGF-I、IGF-II、bFGF、TGF-β1、LIF 和 GM-CSF 等重组生长因子和细胞因子的组合，对体外生产的牛胚胎的发育效果与 10%胎牛血清相似。这种完全合成的胚胎培养方法在胚胎转移的生物安全方面具有不可否认的优势。
