Hanna Carol, Long Charles, Hinrichs Katrin, Westhusin Mark, Kraemer Duane
College of Veterinary Medicine, Department of Physiology and Pharmacology, Texas A&M University, College Station, TX, 77843-4466, USA.
Anim Reprod Sci. 2008 May;105(3-4):451-6. doi: 10.1016/j.anireprosci.2007.12.009. Epub 2007 Dec 28.
To improve assisted reproductive technologies in the domestic dog, different transport treatments were evaluated for their ability to maintain viability of canine oocytes, as assessed by esterase activity 8h after storage or after 48 h of in vitro maturation (IVM) culture. In Experiment 1, ovaries were transported within reproductive tracts or were excised and stored at either 20 or 37 degrees C in phosphate buffered saline. Oocytes collected from reproductive tracts transported at 37 degrees C had the greatest viability after storage (P<0.05). However, after IVM there were no significant differences among any of the four storage conditions in oocyte viability or meiotic resumption (P=0.05). In Experiment 2, isolated oocytes were transported in either TCM-199 with Hank's salts and Hepes buffer or in TL-Hepes at either 20 or 37 degrees C, or in maturation medium equilibrated with 5% CO(2) at 37 degrees C. In Experiment 2, oocytes transported in Hepes buffered media at 37 degrees C had greater viability rates after storage than did those transported in these same media at 20 degrees C or in sodium bicarbonate buffered medium at 37 degrees C (P<0.001). After IVM, oocytes transported in the 37 degrees C treatment groups had greater viability rates than did those transported at 20 degrees C (P<0.01). Overall, isolated oocytes transported at 37 degrees C had greater rates of meiotic resumption than did those transported at 20 degrees C (P<0.05). Taken together, these data indicate that canine oocytes exhibited sensitivity to lesser temperatures and maintained greater rates of viability during transport at 37 degrees C. Isolated oocytes maintained greater viability than oocytes transported in situ. Hepes buffered media increased viability rates for isolated oocytes transported at 37 degrees C compared to a similar medium buffered with sodium bicarbonate.
为改进家犬的辅助生殖技术，对不同的运输处理方式维持犬卵母细胞活力的能力进行了评估，活力通过储存8小时后或体外成熟（IVM）培养48小时后的酯酶活性来评估。在实验1中，卵巢在生殖道内运输，或被切除后在20℃或37℃的磷酸盐缓冲盐溶液中储存。从37℃运输的生殖道中收集的卵母细胞在储存后具有最高的活力（P<0.05）。然而，IVM后，四种储存条件中的任何一种在卵母细胞活力或减数分裂恢复方面均无显著差异（P=0.05）。在实验2中，分离的卵母细胞在含有汉克斯盐和赫佩斯缓冲液的TCM-199中，或在20℃或37℃的TL-赫佩斯中，或在37℃用5%二氧化碳平衡的成熟培养基中运输。在实验2中，在37℃的赫佩斯缓冲培养基中运输的卵母细胞在储存后的活力率高于在相同培养基中20℃运输的卵母细胞或在37℃的碳酸氢钠缓冲培养基中运输的卵母细胞（P<0.001）。IVM后，在37℃处理组中运输的卵母细胞的活力率高于在20℃运输的卵母细胞（P<0.01）。总体而言，在37℃运输的分离卵母细胞的减数分裂恢复率高于在20℃运输的卵母细胞（P<0.05）。综上所述，这些数据表明犬卵母细胞对较低温度敏感，在37℃运输期间保持较高的活力率。分离的卵母细胞比原位运输的卵母细胞保持更高的活力。与用碳酸氢钠缓冲的类似培养基相比，赫佩斯缓冲培养基提高了在37℃运输的分离卵母细胞的活力率。
