摘要
The time course of in vitro red deer nuclear oocyte maturation was determined. Ovaries were obtained at slaughter and oocytes were aspirated from follicles greater than 2 mm in diameter. Oocytes with compact cumulus cells were matured in 50 μl microdrops (10 per drop) under mineral oil containing TCM 199 supplemented with 0.33 mM pyruvate, 10 μg LH and FSH, 1 μg oestradiol and 10% foetal bovine serum. Oocytes were matured at 39 °C and 5% CO2 in air. At 3 h intervals (0–27 h) oocytes were removed from incubation, cumulus expansion scored and removed, and fixed oocytes in ethanol:acetic acid (3:1) for 48 h. Oocytes were stained with lacmoid (1%) and nuclear maturation assessed. Oocytes were arrested in the germinal vesicle (GV) stage at aspiration and up to 6 h of incubation. The nuclear membrane began to disperse after 6 h and by 10.6±0.6 h of incubation 75% of the oocytes exhibited germinal vesicle breakdown (GVBD). The mean time for 50% of the oocytes to reach metaphase one (MI) and metaphase two (MII) was 11.7±0.4 and 24.8±0.9 h, respectively. Cumulus oophorus were tightly compacted at aspiration and did not begin expansion until 12 h of culture. Full expansion was complete by 18 h of culture. Corona radiata cells did not begin expansion until 15 h and were fully expanded by 24 h. Results indicate that in vitro red deer oocyte maturation follows a similar time course of nuclear maturation as reported for bovine and ovine oocytes.
摘要译文
测定体外红鹿核卵母细胞成熟的时间过程。在屠宰时获得卵巢，从直径大于2mm的卵泡吸出卵母细胞。具有致密卵丘细胞的卵母细胞在含有补充有0.33mM丙酮酸盐，10μgLH和FSH的TCM199的矿物油下在50μl微滴（每滴10滴）中成熟，1μg雌二醇和10％胎牛血清。卵母细胞在39℃和5％CO 2空气中成熟。间隔3小时（0-27小时），从孵育中取出卵母细胞，积分扩大计分并除去，并在乙醇：乙酸（3：1）中固定卵母细胞48小时。卵母细胞用lacmoid（1％）染色，评估核成熟。在卵泡发育的囊泡（GV）阶段，在误吸和高达6小时的孵化期间被捕获。 6小时后核膜开始分散，10.6±0。孵育6小时75％的卵母细胞表现出生发泡破裂（GVBD）。 50％卵母细胞达到中期分裂期（MI）和中期分裂期（MII）的平均时间分别为11.7±0.4和24.8±0.9 h，分别。积雨云在吸气时紧紧压缩，直到培养12小时才开始膨胀。 18小时的文化完全扩张完成。电晕辐射细胞直到15小时才开始膨胀，并在24小时内完全膨胀。结果表明体外红鹿卵母细胞成熟遵循与牛和羊卵母细胞相似的核成熟的相似时程。
D.KBerg[a][c];J.GThompson[a][1];G.WAsher[b];. Development of in vitro embryo production systems for red deer (Cervus elaphus)[J]. Animal Reproduction Science, 2002,70(1-2): 77-84