Chen Zhilong, Zhang Caiyong, Yan Chao, Peng Cuiting, Xie Hao, Pan Yingting, Qi Lin, Qu Chaoqun, Zhao Yulan, Tang Zhonglin
Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Key Laboratory of Livestock and Poultry Multi-omics of MARA, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, 518124, China.
Kunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute, Chinese Academy of Agricultural Sciences, Foshan 528226, China.
Anim Reprod Sci. 2025 Aug;279:107945. doi: 10.1016/j.anireprosci.2025.107945. Epub 2025 Jun 18.
The intrinsic quality of porcine oocytes is a critical determinant of early embryonic development during in vitro production (IVP). This study investigated the effects of chemically defined in vitro maturation (IVM) systems supplemented with glutamine, GlutaMAX, insulin-transferrin-selenium (ITS), porcine follicular fluid (pFF), citrate, and growth factors (FGF2, LIF, IGF1, termed FLI) on the cumulus diffusion and maturation of oocytes derived from different diameters follicles. Moreover, four IVM media were systematically assessed for their effects on oocyte nuclear and cytoplasmic maturation and subsequent embryonic development. Results showed that the addition of pFF or FLI alone significantly enhanced cumulus expansion (P < 0.05). FLI medium combined with ITS, citrate, and pFF, exhibited higher average cumulus diffusion diameters and rates of first polar body extrusion. Notably, the synergistic combination of ITS, citrate, and pFF with FLI medium upregulated expression of cumulus expansion makers (HAS2, PTX3, CX43, TNFAIP6) and oocyte maturation makers (GDF9 and BMP15), improved cortical granule distribution, spindle morphology, chromosome alignment, and elevated glutathione (GSH) levels. This combination also enhanced cleavage and blastocyst rates, increased inner cell mass cell counts in blastocysts, and improved enucleation and fusion efficiency of oocytes following somatic cell nuclear transfer (SCNT). Collectively, the combination of FLI medium with ITS, pFF, and citrate synergistically promoted the maturation and developmental potential of porcine oocytes derived from prepubertal gilts, including those from small follicles. This optimized IVM system provides a robust foundation for advancing porcine IVP in reproductive biotechnology.
猪卵母细胞的内在质量是体外生产（IVP）过程中早期胚胎发育的关键决定因素。本研究调查了添加谷氨酰胺、GlutaMAX、胰岛素-转铁蛋白-硒（ITS）、猪卵泡液（pFF）、柠檬酸盐和生长因子（FGF2、LIF、IGF1，称为FLI）的化学成分明确的体外成熟（IVM）系统对不同直径卵泡来源的卵母细胞的卵丘扩散和成熟的影响。此外，系统评估了四种IVM培养基对卵母细胞核成熟和细胞质成熟以及随后胚胎发育的影响。结果表明，单独添加pFF或FLI可显著增强卵丘扩展（P<0.05）。FLI培养基与ITS、柠檬酸盐和pFF组合，表现出更高的平均卵丘扩散直径和第一极体排出率。值得注意的是，ITS、柠檬酸盐和pFF与FLI培养基的协同组合上调了卵丘扩展标志物（HAS2、PTX3、CX43、TNFAIP6）和卵母细胞成熟标志物（GDF9和BMP15）的表达，改善了皮质颗粒分布、纺锤体形态、染色体排列，并提高了谷胱甘肽（GSH）水平。这种组合还提高了卵裂率和囊胚率，增加了囊胚内细胞团细胞数量，并提高了体细胞核移植（SCNT）后卵母细胞的去核和融合效率。总体而言，FLI培养基与ITS、pFF和柠檬酸盐的组合协同促进了来自青春期前后备母猪（包括来自小卵泡的母猪）的猪卵母细胞的成熟和发育潜力。这种优化的IVM系统为推进生殖生物技术中的猪IVP提供了坚实的基础。
