Cordova A, Perreau C, Uzbekova S, Ponsart C, Locatelli Y, Mermillod P
Institut National de Recherche Agronomique (INRA), UMR7247, Physiologie de la Reproduction et des Comportements, Nouzilly, France; Union Nationale des Coopératives d'Elevage et d'Insémination Animale (UNCEIA), Maisons-Alfort, France.
Institut National de Recherche Agronomique (INRA), UMR7247, Physiologie de la Reproduction et des Comportements, Nouzilly, France.
Theriogenology. 2014 Jun;81(9):1163-73. doi: 10.1016/j.theriogenology.2014.01.012. Epub 2014 Jan 31.
The use of somatic cells for coculture with embryos has been amply investigated to study embryo maternal interactions. The use of bovine oviduct epithelial cells (BOEC) has been shown to improve the blastocyst rate and quality, affecting their gene expression profile. In this study, we evaluated different timings of BOEC coculture for the development of in-vitro-produced embryos and their effects on blastocysts rate and mRNA abundance of some genes that are important for embryo development. Our results confirmed the positive effects of BOEC on early development of bovine embryos. The presence of the cells during the first four days or during the last four days of development was enough to produce the full BOEC effect. When the presence of BOEC was restricted to the four first days, the kinetics of blastocyst development was accelerated, with significantly more blastocysts at Days 6 and 7 than when the cells were present all along the culture or only during the last four days. Older cells used at early stage were not active anymore. Using young cells at late stage did not improve the cell effect, compared with the older ones. Therefore, the lower effect of BOEC at late stage, compared with early period, may not be explained by cell aging. In addition, the presence of BOEC, at early or late stages, induced changes in the embryos expression profile of genes known to be related to embryo quality, suggesting reduced apoptosis and increased capacity to struggle against oxidative stress after coculture. In conclusion, we confirmed the effect of BOEC on the rate and quality of bovine IVP embryos development. We found for the first time that the presence of BOEC during the four first days of the 8-days development is enough to produce these effects. These first four days represent the period of the presence of the embryos in the oviduct in vivo, highlighting the physiological relevance of this in vitro model of coculture. In addition, we found that the presence of BOEC at early stages of development induced modification of transcription profile in the blastocyst, four days later, suggesting an epigenetic regulation induced by BOEC in growing embryos.
为研究胚胎与母体之间的相互作用，人们对体细胞与胚胎共培养的应用进行了大量研究。已证实，使用牛输卵管上皮细胞（BOEC）可提高囊胚率和质量，并影响其基因表达谱。在本研究中，我们评估了BOEC共培养的不同时间对体外生产胚胎发育的影响，以及其对囊胚率和一些对胚胎发育重要基因的mRNA丰度的影响。我们的结果证实了BOEC对牛胚胎早期发育具有积极作用。在发育的前四天或后四天存在这些细胞就足以产生完整的BOEC效应。当BOEC的存在仅限于前四天时，囊胚发育的动力学加快，在第6天和第7天的囊胚数量显著多于细胞在整个培养过程中存在或仅在最后四天存在的情况。早期使用的老化细胞不再具有活性。与老化细胞相比，后期使用年轻细胞并未改善细胞效应。因此，与早期相比，BOEC在后期的作用较低可能无法用细胞老化来解释。此外，BOEC在早期或晚期的存在会诱导胚胎中已知与胚胎质量相关基因的表达谱发生变化，这表明共培养后细胞凋亡减少，抵抗氧化应激的能力增强。总之，我们证实了BOEC对牛体外生产胚胎发育的速率和质量具有影响。我们首次发现，在8天发育的前四天存在BOEC就足以产生这些效应。这前四天代表了胚胎在体内输卵管中的存在时期，突出了这种体外共培养模型的生理相关性。此外，我们发现BOEC在发育早期的存在会在四天后诱导囊胚转录谱的改变，这表明BOEC在生长中的胚胎中诱导了表观遗传调控。
