Matoba Satoko, Fair Trudee, Lonergan Patrick
National Livestock Breeding Center, Odakurahara 1, Nishigo, Fukushima 961-8511, Japan.
Reprod Fertil Dev. 2010;22(5):839-51. doi: 10.1071/RD09277.
The ability to successfully culture oocytes and embryos individually would facilitate the study of the relationship between follicle parameters and oocyte developmental competence, in order to identify markers of competent oocytes, as well as the ability to use small numbers of oocytes from an individual donor such as when ovum pick-up is carried out. Using a total of 3118 oocytes, the aim of the present study was to develop a system capable of supporting the development of immature bovine oocytes to the blastocyst stage in an individually identifiable manner. Initially, post-fertilisation embryo culture in the Well-of-the-Well (WOW) system, on the cell adhesive Cell-Tak or in polyester mesh was tested and shown to result in similar development to embryos cultured in standard group culture. The results demonstrate that it is possible to culture bovine oocytes to the blastocyst stage in an individually identifiable manner in all three culture systems with comparable success rates. This permits the localisation and identification of individual embryos throughout preimplantation development in vitro while retaining the developmental benefits of group culture. In terms of ease of preparation and use, culture in isolation within the strands of a polyester mesh is preferable.
成功地对卵母细胞和胚胎进行单独培养,将有助于研究卵泡参数与卵母细胞发育能力之间的关系,以确定有发育能力的卵母细胞的标志物,以及能够使用来自单个供体的少量卵母细胞,例如在进行采卵时。本研究共使用3118个卵母细胞,目的是开发一种能够以个体可识别的方式支持未成熟牛卵母细胞发育到囊胚阶段的系统。最初,对在“孔中孔”(WOW)系统中、在细胞黏附剂Cell-Tak上或在聚酯网中进行受精后胚胎培养进行了测试,结果表明其发育情况与在标准群体培养中培养的胚胎相似。结果表明,在所有三种培养系统中,都有可能以个体可识别的方式将牛卵母细胞培养到囊胚阶段,成功率相当。这使得在体外整个植入前发育过程中能够定位和识别单个胚胎,同时保留群体培养的发育优势。就制备和使用的简便性而言,在聚酯网的股线内进行单独培养更为可取。