Constantine the Philosopher University in Nitra, Nitra, Slovakia.
Institute of Animal Physiology and Genetics, Czech Academy of Sciences, Liběchov, Czech Republic.
Sci Rep. 2024 Mar 25;14(1):7081. doi: 10.1038/s41598-024-57865-7.
In this article, we focused on the impact of precisely chemically modified FLI maturation medium enriched with fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), insulin-like growth factor 1 (IGF1), and polyvinyl alcohol (PVA) and its potential to improve the efficiency of in vitro production of porcine embryos. We hypothesized that enhancing the composition of the maturation medium could result in an elevated production of embryos in vitro and can affect EGA. FLI medium resulted in a significantly higher rate of oocyte blastocyst maturation and formation compared to the control DMEM medium. In addition, immunocytochemical labelling confirmed the detection of UBF in 4-cell FLI parthenogenic embryos, suggesting similarities with natural embryo development. Through RNAseq analysis, upregulated genes present in 4-cell FLI embryos were found to play key roles in important biological processes such as cell proliferation, cell differentiation, and transcriptional regulation. Based on our findings, we demonstrated the positive influence of FLI medium in the evaluation of in vitro embryo production, EGA detection, transcriptomic and proteomic profile, which was confirmed by the positive activation of the embryonal genome in the 4-cell stage of parthenogenetically activated embryos.
本文重点研究了经精心化学修饰的富含成纤维细胞生长因子 2(FGF2)、白血病抑制因子(LIF)、胰岛素样生长因子 1(IGF1)和聚乙烯醇(PVA)的 FLI 成熟培养基对体外生产猪胚胎效率的影响及其潜在作用。我们假设,增强成熟培养基的组成可能会提高体外胚胎的产量,并可能影响胚胎基因组激活(EGA)。与对照 DMEM 培养基相比,FLI 培养基显著提高了卵母细胞囊胚成熟和形成的比率。此外,免疫细胞化学标记证实了在 4 细胞 FLI 孤雌胚胎中检测到 UBF,表明与自然胚胎发育具有相似性。通过 RNAseq 分析,发现 4 细胞 FLI 胚胎中上调的基因在细胞增殖、细胞分化和转录调控等重要生物学过程中发挥关键作用。基于我们的发现,我们证明了 FLI 培养基在体外胚胎生产、EGA 检测、转录组和蛋白质组谱评估中的积极影响,这通过孤雌激活胚胎的 4 细胞阶段胚胎基因组的阳性激活得到了证实。