权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of a transient co-incubation IVF system in the pig

猪瞬时共孵化 IVF 系统的开发

基本信息

批准号：
16580230
负责人：
FUNAHASHI Hiroaki
金额：
$ 2.3万
依托单位：
OKAYAMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580230/
关键词：
Pigs In vitro fertilization Polyspermy Caffeine Oxidative stress Sperm capacitation Acrosome reaction 共培養

项目摘要

This research project was undertaken to develop a transient-co-incubation IVF system in the pig, in order to obtain an efficient normal monospermic sperm penetration. Obtained sights are as follows :(A)Effect of pretreatment of spermatozoa with adenosine and a transient gamete co-incubation with caffeineA new transient co-incubation IVF system, in which denuded oocytes are co-cultured with spermatozoa in medium containing caffeine for 5 to 30 min and then continuing the culture in caffeine-free medium, will reduce the incidence of polyspermic penetration. Preincubation of fresh spermatozoa with adenosine before the transient co-incubation IVF can also improve the monospermic penetration.(B)Effective regulation of sperm capacitation during transient IVF periodThe presence of beta-mercaptoethanol (bME) prevents induction of sperm capacitation even in the presence of caffeine, whereas H_2O_2 induces sperm capacitation and the spontaneous acrosome reaction.(C)Regulation of oxidative stress of gametes during transient co-incubationSupplementation with bME during IVF procedures, except during a transient co-culture period of gametes in the presence of caffeine, has a beneficial effect in maintaining the function of gametes, the incidence of normal fertilization and, consequently, the quality of IVF embryos.(D)Selection of antioxidants to improve the function of extended boar semen stored at a low temperatureGlutathione and cysteine can improve the viability and functional status of boar spermatozoa during liquid preservation and boar spermatozoa penetrated in vitro even after preservation in the presence of cysteine at 10℃ for 29 days.(E)Development of an IVM system for the transient IVF systemAlthough cytoplasmic maturation in porcine oocytes pretreated with roscovitine for 48 h did not equal that of control oocytes in the current IVM system, those stored oocytes can matured, fertilized and developed in vitro to the blastocyst stage.

该研究项目旨在开发猪体内的瞬时共孵化体外受精系统，以获得有效的正常单精子穿透能力。获得的结果如下：(A)用腺苷预处理精子和用咖啡因瞬时共孵育配子的效果一种新的瞬时共孵育IVF系统，其中将裸卵母细胞与精子在含咖啡因的培养基中共培养5至30分钟，然后在无咖啡因的培养基中继续培养，将降低发生率多精渗透。在短暂共孵育 IVF 之前将新鲜精子与腺苷预孵育也可以提高单精子穿透力。(B)短暂 IVF 期间精子获能的有效调节β-巯基乙醇 (bME) 的存在即使在存在咖啡因的情况下也能阻止精子获能的诱导，而 H_2O_2 会诱导精子获能并导致精子获能。 (C) 瞬时共孵育期间配子氧化应激的调节除了在存在咖啡因的情况下配子的瞬时共培养期间外，在 IVF 过程中补充 bME 对于维持配子功能、正常受精的发生率以及 IVF 胚胎的质量具有有益的作用。(D) 选择抗氧化剂改善低温保存的延长公猪精液的功能谷胱甘肽和半胱氨酸可以改善公猪精子在液体保存过程中的活力和功能状态，并且即使在10℃半胱氨酸存在下保存29天后，公猪精子仍能在体外渗透。（E）用于瞬时IVF系统的IVM系统的开发用roscovitine预处理48 h的猪卵母细胞与当前IVM系统中的对照卵母细胞不相等，这些储存的卵母细胞可以在体外成熟、受精并发育至囊胚阶段。