College of Animal Science and Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China.
Theriogenology. 2011 Sep 1;76(4):700-4. doi: 10.1016/j.theriogenology.2010.10.039. Epub 2011 May 23.
Three media were evaluated for their ability to support in vitro maturation of donkey (Equus asinus) oocytes and their development after parthenogenetic activation. The basal medium for Medium 1 (M1) and Medium 2 (M2) was M199 and DMEM/F12 respectively, whereas, Medium 3 (M3) consisted of equal parts (v/v) of M199 and DMEM/F12. All three media were supplemented with 10% (v/v) fetal calf serum, 0.01 units/mL porcine FSH, 0.01 units/mL equine LH, 200 ng/mL insulin-like growth factor 1(IGF-I), 10 μl/mL insulin-transferrin-selenium (ITS), 0.1 mg/mL taurine, 0.1 mg/mL L-cysteine, 0.05 mg/mL L-glutamine, 0.11 mg/mL sodium pyruvate, and 25 mg/mL gentamycin. There were no significant differences among the three maturation media for oocyte maturation. Maturation rate of donkey oocytes in M1 was 53% for compact (Cp) cumulus-oocyte complexes and 75% for expanded (Ex) cumulus-oocyte complexes; in M2 these were 55 and 77%, respectively; and in M3, 58 and 75%. The percentage of cleaved parthenotes and 4- or 8-cell embryos were not significantly different for oocytes matured in the various media (61 and 24% for M1; 66 and 32% for M2; and 67 and 33% for M3). Oocytes matured in M3 tended to yield a higher rate of advanced embryo development (morula) than oocytes matured in M1 (22 vs 9%; P = 0.07). In conclusion, donkey oocytes were matured and parthenogenetically activated in vitro, using methods similar to those used in the horse.
三种培养基用于支持驴卵母细胞体外成熟及其孤雌激活后发育的能力评估。培养基 1(M1)和培养基 2(M2)的基础培养基分别为 M199 和 DMEM/F12,而培养基 3(M3)由 M199 和 DMEM/F12 等体积(v/v)组成。这三种培养基均添加了 10%(v/v)胎牛血清、0.01 单位/mL 猪 FSH、0.01 单位/mL 马 LH、200ng/mL 胰岛素样生长因子 1(IGF-1)、10μL/mL 胰岛素转铁蛋白-硒(ITS)、0.1mg/mL 牛磺酸、0.1mg/mL L-半胱氨酸、0.05mg/mL L-谷氨酰胺、0.11mg/mL 丙酮酸钠和 25mg/mL 庆大霉素。三种成熟培养基对卵母细胞成熟没有显著差异。M1 中致密(Cp)卵丘-卵母细胞复合物的卵母细胞成熟率为 53%,扩张(Ex)卵丘-卵母细胞复合物为 75%;在 M2 中分别为 55%和 77%;在 M3 中,分别为 58%和 75%。在不同培养基中成熟的卵母细胞的卵裂孤雌胚和 4-或 8-细胞胚胎的百分比没有显著差异(M1 为 61%和 24%;M2 为 66%和 32%;M3 为 67%和 33%)。在 M3 中成熟的卵母细胞在体外发育为高级胚胎(桑葚胚)的比例高于在 M1 中成熟的卵母细胞(22 对 9%;P=0.07)。总之,驴卵母细胞在体外使用类似于马的方法进行成熟和孤雌激活。