Smetanina I G, Tatarinova L V, Krivokharchenko A S
All-Russian Institute of Physiology, Biochemistry, and Nutrition of Farm Animals, Russian Academy of Agricultural Sciences, Borovsk, Kaluga oblast, Russia.
Ontogenez. 2000 Mar-Apr;31(2):139-43.
We studied the capacity of the cattle oocyte for the resumption of meiosis and achievement of metaphase II in various protein-free culture media (DMEM, TCM-199, Ham's F-10, and Ham's F-12) and the pattern of influence of the estrous serum on in vitro development of fertilized cattle oocytes, with special reference to the time of its addition to the synthetic oviduct fluid containing BSA. In the first experimental series, it was shown that the highest number of oocytes (76.1%) resumed meiosis in DMEM medium. Meiosis was not resumed in Ham's F-12. Intermediate results were obtained for TCM-199 (55.1%), which is commonly used for maturation of cattle oocytes in vitro, and for Ham's F-10 (51.7%). The oocytes reached metaphase II in DMEM at a higher rate (45.3%) than in TCM-199 or Ham's F-10 (29.4 and 8.6%, respectively). In the second experimental series, the estrous serum was added to the culture medium within 20 h (control) or 42 h (experiment) after the beginning of fertilization. The estrous serum did not inhibit the first cleavage division (the percentage of cleaving embryos did not differ reliably: 32.7 and 37.9%, respectively). However, a later serum addition to the culture medium (within 42 h after the beginning of fertilization) reliably increased the percentage of embryos that reached the blastocyst stage (6.5% in the control and 17.8% in the experiment) and the hatched blastocyst stage (2 and 9.2%, respectively).
我们研究了牛卵母细胞在各种无蛋白培养基（DMEM、TCM - 199、Ham's F - 10和Ham's F - 12）中恢复减数分裂并达到中期II的能力，以及发情期血清对体外受精牛卵母细胞发育的影响模式，特别参考了其添加到含牛血清白蛋白的合成输卵管液中的时间。在第一个实验系列中，结果表明，在DMEM培养基中，恢复减数分裂的卵母细胞数量最多（76.1%）。在Ham's F - 12中卵母细胞未恢复减数分裂。对于常用于牛卵母细胞体外成熟的TCM - 199（55.1%）和Ham's F - 10（51.7%），得到的是中间结果。卵母细胞在DMEM中达到中期II的比例（45.3%）高于在TCM - 199或Ham's F - 10中的比例（分别为29.4%和8.6%）。在第二个实验系列中，在受精开始后的20小时内（对照组）或42小时内（实验组）将发情期血清添加到培养基中。发情期血清未抑制第一次卵裂（分裂胚胎的百分比无显著差异：分别为32.7%和37.9%）。然而，在受精开始后较晚时间（42小时内）向培养基中添加血清，确实增加了达到囊胚阶段的胚胎百分比（对照组为6.5%，实验组为17.8%）和孵化囊胚阶段的胚胎百分比（分别为2%和9.2%）。
