Whitty Annie L, Kind Karen L, Dunning Kylie R, McPherson Nicole O, Nottle Mark B
Robinson Research Institute, The University of Adelaide, South Australia, Australia; Discipline of Reproduction and Development, School of Biomedicine, The University of Adelaide, South Australia, Australia.
Robinson Research Institute, The University of Adelaide, South Australia, Australia; Discipline of Reproduction and Development, School of Biomedicine, The University of Adelaide, South Australia, Australia; Davies Livestock Research Centre, School of Animal and Veterinary Sciences, The University of Adelaide, South Australia, Australia.
Theriogenology. 2025 Mar 15;235:46-55. doi: 10.1016/j.theriogenology.2024.12.022. Epub 2025 Jan 2.
In vitro embryo production (IVP) is used in the cattle industry to increase the rate of genetic gain. IVP uses semen that has been frozen and thawed, a process that renders sperm less viable than sperm from fresh semen. Granulocyte macrophage colony stimulating factor (GM-CSF) is present in bovine seminal plasma, while its receptor is present on bovine sperm. The present study aimed to determine if GM-CSF could improve the function and quality of frozen-thawed bovine sperm and embryo development following in vitro fertilization. Thawed bovine sperm (n = 3 bulls with 3 replicates per bull) was incubated with 0, 0.1, 1, 2 or 10 ng/ml of recombinant bovine GM-CSF in buffered wash media for 45 min and assessed for motility, glucose uptake, mitochondrial activity, intracellular calcium, capacitation, DNA integrity, and in vitro embryo development. The addition of 1, 2, and 10 ng/ml GM-CSF increased total motility (P = 0.02, P = 0.007, P = 0.01), progressive (P = 0.02, P = 0.03, P = 0.01), and rapid motility (P = 0.01, P = 0.01, P = 0.01), while 10 ng/ml increased glucose uptake (P = 0.003), and 1, 2, and 10 ng/ml increased capacitation (P = 0.003, P = 0.001, P = 0.0003). There was no difference between groups for mitochondrial activity, intracellular calcium, or DNA integrity. GM-CSF treatment of sperm prior to in vitro insemination increased fertilization rate (P = 0.01), hatching blastocyst rate (P = 0.05), and blastocyst inner cell mass cell number (P = 0.03) compared with control. In conclusion, GM-CSF treatment of frozen-thawed bovine sperm improves sperm function and quality resulting in increased fertilization capacity and subsequent embryo development, suggesting it may improve cattle IVP efficiencies.
体外胚胎生产（IVP）在养牛业中用于提高遗传增益率。IVP使用经过冷冻和解冻的精液，这一过程会使精子的活力低于新鲜精液中的精子。粒细胞巨噬细胞集落刺激因子（GM-CSF）存在于牛精液中，而其受体存在于牛精子上。本研究旨在确定GM-CSF是否能改善冷冻解冻牛精子的功能和质量以及体外受精后的胚胎发育。将解冻的牛精子（n = 3头公牛，每头公牛3个重复）在缓冲洗涤培养基中与0、0.1、1、2或10 ng/ml的重组牛GM-CSF孵育45分钟，并评估其活力、葡萄糖摄取、线粒体活性、细胞内钙、获能、DNA完整性和体外胚胎发育情况。添加1、2和10 ng/ml的GM-CSF可提高总活力（P = 0.02，P = 0.007，P = 0.01）、前进活力（P = 0.02，P = 0.03，P = 0.01）和快速活力（P = 0.01，P = 0.01，P = 0.01），而10 ng/ml可提高葡萄糖摄取（P = 0.003），1、2和10 ng/ml可提高获能（P = 0.003，P = 0.001，P = 0.0003）。各组之间在线粒体活性、细胞内钙或DNA完整性方面没有差异。与对照组相比，体外受精前对精子进行GM-CSF处理可提高受精率（P = 0.01）、囊胚孵化率（P = 0.05）和囊胚内细胞团细胞数（P = 0.03）。总之，对冷冻解冻的牛精子进行GM-CSF处理可改善精子功能和质量，从而提高受精能力和随后的胚胎发育，表明它可能提高牛IVP效率。
