Studies on production of pig embryos from growing oocytes

利用生长中的卵母细胞生产猪胚胎的研究

基本信息

批准号：
09556061
负责人：
KATO Seishiro
金额：
$ 9.09万
依托单位：
KOBE UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09556061/
关键词：
pig oogenesis folliculogenesis oocyte growth oocyte maturation in vitro culture in vitro fertilization embryo production ブタ体外発育卵母細胞体外成熟卵母細胞

项目摘要

Several hundred thausand primordial follicles are contained in the ovaries of domestic animals. The development of in vitro culture systems for the growth of small oocytes in primordial or small follicles is expected to provide a new source of a large population of oocytes for livestock production. This study was conducted to develop Culture systems for small oocytes from pig ovaries.Preantral follicles containing oocytes with various sizes were isolated from pig ovaries and cultured in collagen gel for up to 16 days in the presence of serum, FSH and oestradiol. Waymouth MB7 52/1 was used as a basic culture medium for oocyte growth. After culture, the oocytes enclosed by one or two layers of cumulus cells were cultured for meiotic maturation and inseminated with spermatozoa. The results obtained were as follows.1. Pig growing oocytes grew to their final size, acquired meiotic competence du ring in vitro culture, and were penetrated by spcermatozoa in vitro, although there was a very lo … More w level of success in fertilization. In contrast to the growing oocytes, no culture methods were available for much smaller oocytes. We tried to culture primordial follicles which contained small oocytes 30 μm in diameter, but the integrity of the follicles was soon broken and the follicles survived for only a few days.2. Using culture systems, we demonstrated that granulosa cells regulate oocyte growth, and that antrum formation of granulosa cells was induced only by meiotic-competent oocytes, indicating that the oocytes secrete a substance(s) that induces differentiation of granulosa cells.3. Of four glycosaminoglycans examined, hyalronic acids and chondroitin sulfate A supported the development of in vitro-matured and -fertilized pig oocytes to the blastocyst stage.4. In addition, degradation of cyclin B1 molecules and MAP kinase deph osphorylation during fertilization, localisation of phosphorylated MAP kinase during the transition from meiosis I to meiosis II, assembly and transformation of the spindle during the progression through meiotic cell cycle, and the level and localization of CENP-E and kinetochore numaber during maturation were examined. Less

houndred thausand thaus和primordialz含有家畜的卵巢中。在血清，FSH和Oestradiol的存在下，有多种含量从卵巢中进行的，在胶原蛋白凝胶中培养长达16天。用于成熟的培养物和精子的培养，所获得的结果如下1。卵母细胞的生长，没有培养的卵母细胞，我们试图培养培养较小的卵母细胞30μm的原始培养物，而卵泡仅存活了几天。卵母细胞的生长以及颗粒细胞的饱和形成是诱导颗粒细胞区分的卵母细胞。3。 .4。除了受精期间的细胞周期蛋白B1分子和MAP DEPH脱磷，在从减数分裂I到减数分裂II的过渡期间，磷酸化的MAP激酶的定位，进度细胞周期的组装和转化，以及CENP---末端的动力学数量较少