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在猪卵母细胞体外成熟过程中,在特定培养基中添加转化生长因子-α可提高其发育能力和细胞内超微结构。

Transforming growth factor-alpha in a defined medium during in vitro maturation of porcine oocytes improves their developmental competence and intracellular ultrastructure.

机构信息

Research Institute for the Functional Peptides, Shimojo, Yamagata, Japan.

出版信息

Theriogenology. 2009 Oct 1;72(6):841-50. doi: 10.1016/j.theriogenology.2009.06.004. Epub 2009 Jul 25.

Abstract

We examined the effects of transforming growth factor-alpha (TGF-alpha) to develop a defined medium for in vitro maturation (IVM) of porcine (Sus scrofa domesticus) oocytes. Cumulus-oocyte complexes (COCs) were matured in porcine oocyte medium containing 3mg/mL polyvinyl alcohol (POM) and TGF-alpha (0, 1, 10, or 100 ng/mL) in the presence or absence of the gonadotropins equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG). In the absence of gonadotropins, adding 10 ng/mL TGF-alpha increased (P<0.05) the percentage of oocytes that reached metaphase II (24.2%) compared with that of the control (no TGF-alpha addition; 5.6%). In the presence of gonadotropins, although maturation rate did not differ among TGF-alpha treatments (75.4% to 84.8%), the rate of blastocyst formation (28.1%) was higher (P<0.05) in the TGF-alpha group (28.1%) than that in the control group (15.9%) after in vitro fertilization and embryo culture. An electron microscope study revealed that TGF-alpha-treated oocytes contained more homogenous lipid droplets than did control oocytes. Moreover, mitochondria surrounded by the endoplasmic reticulum were observed only in the TGF-alpha-treated oocytes. In blastocysts derived from the latter oocytes, mitochondria with numerous cristae were frequently observed compared with that in blastocysts from control oocytes. When the Day-5 blastocysts obtained from oocytes matured with TGF-alpha were surgically transferred into four recipients, a total of 29 piglets were farrowed. We concluded that the addition of TGF-alpha to the defined IVM medium of porcine oocytes improved the subsequent blastocyst formation and that the blastocysts produced by the defined in vitro production system have developmental competence to full term after embryo transfer.

摘要

我们研究了转化生长因子-α(TGF-α)对建立猪卵母细胞体外成熟(IVM)定义培养基的影响。在含有 3mg/mL 聚乙烯醇(POM)的猪卵母细胞培养基中,添加或不添加促性腺激素马绒毛膜促性腺激素(eCG)和人绒毛膜促性腺激素(hCG),使卵丘-卵母细胞复合物(COCs)成熟。在没有促性腺激素的情况下,添加 10ng/mL TGF-α可增加(P<0.05)达到中期 II 期的卵母细胞百分比(24.2%),而对照组(无 TGF-α添加;5.6%)。在有促性腺激素的情况下,尽管 TGF-α处理之间的成熟率没有差异(75.4%至 84.8%),但在体外受精和胚胎培养后,囊胚形成率(28.1%)在 TGF-α组(28.1%)高于对照组(15.9%)。电镜研究表明,TGF-α处理的卵母细胞比对照组卵母细胞含有更多均匀的脂滴。此外,只有在 TGF-α处理的卵母细胞中才观察到被内质网包围的线粒体。在来自后者卵母细胞的囊胚中,与来自对照组卵母细胞的囊胚相比,经常观察到具有许多嵴的线粒体。当将用 TGF-α成熟的卵母细胞获得的 Day-5 囊胚通过手术移植到四个受体中时,总共分娩了 29 头小猪。我们得出结论,向猪卵母细胞的定义 IVM 培养基中添加 TGF-α可提高随后的囊胚形成率,并且通过定义的体外生产系统产生的囊胚在胚胎移植后具有发育到足月的能力。

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