Ocampo M B, Ocampo L C, Mori T, Ueda J, Kanagawa H
Department of Theriogenology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
J Vet Med Sci. 1994 Feb;56(1):173-6. doi: 10.1292/jvms.56.173.
Studies were conducted to determine if the amniotic fluid of a developing chick embryo (CEAm) could be used as a co-culture system for the maturation of pig oocytes. In Exp. 1, the pig oocytes were cultured in mTCM-199 (control) or day 3, 4 and 5 CEAm. The maturation rate of pig oocytes in the control vs CEAm was significantly higher (P < 0.001). The maturation rate of pig oocytes cultured in day 3 vs day 4 and 5 CEAm was significantly higher (P < 0.001). In Exp. 2, the pig oocytes were cultured in day 3 CEAm maintained either in a conventional incubator with 40-70% relative humidity or incubator with 5% CO2 in air atmosphere at 37 degrees C or 39 degrees C. A significantly lower maturation rate (P < 0.001) was obtained when CEAm were maintained in an incubator with 5% CO2 gas atmosphere regardless of the temperature. In Exp. 3, pig oocytes cultured in day 3 CEAm and mTCM-199 (control) were fertilized in vitro (IVF). The penetration and male pronucleus (MPN) formation of pig oocytes cultured in day 3 CEAm was significantly lower (P < 0.01) than those cultured in mTCM-199 medium. The results presented here demonstrate that CEAm could be used to induce maturation of pigs oocytes, however its efficacy is influenced by factors such as developmental stages of chick embryos, temperature and gas atmosphere.
开展了多项研究,以确定发育中的鸡胚羊水(CEAm)是否可用作猪卵母细胞成熟的共培养系统。在实验1中,将猪卵母细胞培养于mTCM - 199(对照)或第3、4和5天的CEAm中。对照组与CEAm组中猪卵母细胞的成熟率显著更高(P < 0.001)。培养于第3天CEAm中的猪卵母细胞的成熟率比培养于第4和5天CEAm中的显著更高(P < 0.001)。在实验2中,将猪卵母细胞培养于第3天的CEAm中,分别置于相对湿度为40 - 70%的传统培养箱或37℃或39℃、含5%二氧化碳的空气环境培养箱中。无论温度如何,当CEAm置于含5%二氧化碳气体环境的培养箱中时,成熟率显著更低(P < 0.001)。在实验3中,将培养于第3天CEAm和mTCM - 199(对照)中的猪卵母细胞进行体外受精(IVF)。培养于第3天CEAm中的猪卵母细胞的穿透率和雄原核(MPN)形成率显著低于培养于mTCM - 199培养基中的(P < 0.01)。此处呈现的结果表明,CEAm可用于诱导猪卵母细胞成熟,然而其效果受鸡胚发育阶段、温度和气体环境等因素影响。