The combination of FLI, ITS, pFF, and citrate improves the developmental potential of gilt oocytes during in vitro maturation

IF 2.2 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science Pub Date : 2025-06-18 DOI:10.1016/j.anireprosci.2025.107945

Zhilong Chen , Caiyong Zhang , Chao Yan , Cuiting Peng , Hao Xie , Yingting Pan , Lin Qi , Chaoqun Qu , Yulan Zhao , Zhonglin Tang

{"title":"The combination of FLI, ITS, pFF, and citrate improves the developmental potential of gilt oocytes during in vitro maturation","authors":"Zhilong Chen , Caiyong Zhang , Chao Yan , Cuiting Peng , Hao Xie , Yingting Pan , Lin Qi , Chaoqun Qu , Yulan Zhao , Zhonglin Tang","doi":"10.1016/j.anireprosci.2025.107945","DOIUrl":null,"url":null,"abstract":"<div><div>The intrinsic quality of porcine oocytes is a critical determinant of early embryonic development during <em>in vitro</em> production (IVP). This study investigated the effects of chemically defined <em>in vitro</em> 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 (<em>P</em> < 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 (<em>HAS2</em>, <em>PTX3</em>, <em>CX43</em>, <em>TNFAIP6</em>) and oocyte maturation makers (<em>GDF9</em> and <em>BMP15</em>), 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.</div></div>","PeriodicalId":7880,"journal":{"name":"Animal Reproduction Science","volume":"279 ","pages":"Article 107945"},"PeriodicalIF":2.2000,"publicationDate":"2025-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Animal Reproduction Science","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0378432025001848","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}

引用次数: 0

Abstract

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.

查看原文本刊更多论文

FLI、ITS、pFF和柠檬酸盐的联合使用提高了后备卵母细胞在体外成熟过程中的发育潜力

猪卵母细胞的内在质量是体外生产（IVP）早期胚胎发育的关键决定因素。本研究研究了添加谷氨酰胺、谷氨酰胺、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）的化学定义体外成熟（IVM）系统对来自不同直径卵泡的卵母细胞积云扩散和成熟的影响。此外，系统地评估了四种IVM培养基对卵母细胞核和细胞质成熟以及随后的胚胎发育的影响。结果表明，单独添加pFF或FLI均能显著增强积云扩张（P <； 0.05）。与ITS、柠檬酸盐和pFF混合的FLI介质表现出更高的平均积云扩散直径和第一极体挤压速率。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云扩增因子（HAS2、PTX3、CX43、TNFAIP6）和卵母细胞成熟因子（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。该组合还提高了卵裂率和囊胚率，增加囊胚内细胞质量细胞计数，并提高了体细胞核移植（SCNT）后卵母细胞的去核和融合效率。总的来说，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的猪卵母细胞的成熟和发育潜力，包括来自小卵泡的卵母细胞。该优化的体外受精系统为推进猪体外受精在生殖生物技术中的应用奠定了坚实的基础。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Animal Reproduction Science 农林科学-奶制品与动物科学

CiteScore

4.50

自引率

9.10%

发文量

136

审稿时长

54 days

期刊介绍： Animal Reproduction Science publishes results from studies relating to reproduction and fertility in animals. This includes both fundamental research and applied studies, including management practices that increase our understanding of the biology and manipulation of reproduction. Manuscripts should go into depth in the mechanisms involved in the research reported, rather than a give a mere description of findings. The focus is on animals that are useful to humans including food- and fibre-producing; companion/recreational; captive; and endangered species including zoo animals, but excluding laboratory animals unless the results of the study provide new information that impacts the basic understanding of the biology or manipulation of reproduction. The journal''s scope includes the study of reproductive physiology and endocrinology, reproductive cycles, natural and artificial control of reproduction, preservation and use of gametes and embryos, pregnancy and parturition, infertility and sterility, diagnostic and therapeutic techniques. The Editorial Board of Animal Reproduction Science has decided not to publish papers in which there is an exclusive examination of the in vitro development of oocytes and embryos; however, there will be consideration of papers that include in vitro studies where the source of the oocytes and/or development of the embryos beyond the blastocyst stage is part of the experimental design.