KOIKE Takayuki, MATSUURA Koji, NARUSE Keiji, FUNAHASHI Hiroaki
Department of Animal Science, Graduate School of Natural Science and Technology, Okayama University, Japan.
J Reprod Dev. 2010 Oct;56(5):552-7. doi: 10.1262/jrd.10-041h. Epub 2010 Jun 16.
Under physiological conditions, mammalian oocytes and embryos appear to be stimulated not only chemically but also mechanically, such as by compression, shear stress and/or friction force in the follicle and female reproductive tract. The present study was undertaken to examine the effects of kinetic culture with a tilting device in chemically defined media during in vitro maturation (IVM) of porcine oocytes and in vitro culture (IVC) following in vitro fertilization (IVF) on the early developmental competence and quality of blastocysts. After culture in a chemically defined IVM medium, modified porcine oocyte medium (mPOM) containing gonadotropins and dibutyryl cAMP for 20 h, the mean diameter of the cumulus-oocyte complexes (COCs) was larger in the tilting culture than in the static controls, whereas the diameter of the oocytes did not differ. When culture of the COCs was continued additionally in a fresh medium without gonadotropins and dibutyryl cAMP for 24 h, the incidences of oocytes completing GVBD and developing to the metaphase-II stage did not differ between the tilting and static culture systems. Furthermore, the sperm penetration after IVF and developmental competence of the oocytes to the blastocyst stage were not different between the tilting and static systems during IVM and IVC. However, tilting culture during both IVM and IVC had a significant positive effect on the number of cells per blastocyst (P<0.05). These observations indicate that tilting culture during IVM and IVC in chemically defined media improves the quality of blastocyst, as determined by the number of cells per blastocyst, without any effects on penetrability and developmental competence.
在生理条件下,哺乳动物的卵母细胞和胚胎似乎不仅受到化学刺激,还受到机械刺激,例如卵泡和雌性生殖道中的压缩、剪切应力和/或摩擦力。本研究旨在探讨在猪卵母细胞体外成熟(IVM)期间以及体外受精(IVF)后的体外培养(IVC)过程中,使用倾斜装置在化学成分明确的培养基中进行动态培养对囊胚早期发育能力和质量的影响。在化学成分明确的IVM培养基(含有促性腺激素和二丁酰环磷腺苷的改良猪卵母细胞培养基,即mPOM)中培养20小时后,倾斜培养组中卵丘-卵母细胞复合体(COCs)的平均直径大于静态对照组,而卵母细胞的直径没有差异。当在不含促性腺激素和二丁酰环磷腺苷的新鲜培养基中继续培养COCs 24小时时,倾斜培养系统和静态培养系统中完成生发泡破裂(GVBD)并发育到中期II期的卵母细胞发生率没有差异。此外,在IVM和IVC期间,倾斜培养系统和静态培养系统在IVF后的精子穿透率以及卵母细胞发育到囊胚阶段的能力方面没有差异。然而,IVM和IVC期间的倾斜培养对每个囊胚的细胞数量有显著的积极影响(P<0.05)。这些观察结果表明,在化学成分明确的培养基中进行IVM和IVC期间的倾斜培养可提高囊胚质量,以每个囊胚的细胞数量来衡量,且对穿透性和发育能力没有任何影响。