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首页> 外文期刊>Small Ruminant Research >In vitro maturation of caprine oocytes in different concentrations of estrous goat serum.
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In vitro maturation of caprine oocytes in different concentrations of estrous goat serum.

机译：不同浓度的发情山羊血清中山羊卵母细胞的体外成熟。

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The aim of the study was to evaluate the optimum concentration of the estrous goat serum for goat in vitro oocyte maturation. Goat ovaries were collected from a local abattoir and transported within 3 h to the laboratory in a warm saline solution (30-35 degrees C), supplemented with 100 IU penicillin G and 100 micro g streptomycin sulfate/ml. Cumulus-oocyte complexes (COC's) were obtained by puncture of 1-4 mm diameter follicles with a 18G needle attached to a disposable syringe, washed 10 times in TCM-199 hepes modification with Earl's salts and supplemented with 50 micro g/ml gentamycin, 100 micro g/ml l-glutamine and 0.25 mM sodium pyruvate without any serum supplementation. The COC's were randomly divided into four groups. Group 1 (n=105) COC's were fresh control and cultured in TCM-199 medium without serum supplementation. Group 2 (n=105) COC's were washed five times and cultured in TCM-199 medium supplemented with 10% EGS. Group 3 (n=110) COC's were washed five times and cultured in TCM-199 medium supplemented with 15% EGS. Group 4 (n=112) COC's were washed five times and cultured in TCM-199 medium supplemented with 20% EGS. After 24-27 h of IVM, oocytes were denuded with the aid of 0.1% hyaluronidase and passing them through a fine pipette, fixed for 24-48 h in a mixture of acetic acid and alcohol (1:3) at room temperature, stained for 10 min with 1% (w/v) orcein in 45% acetic acid and examined for the evidence of different stages of maturation. Significantly higher (p<0.01) maturation rates of oocytes (61-78%) were observed in all concentrations of estrous goat serum compared to those without serum supplementation. However, no significant difference was observed between the 10 and 15% serum supplemented group. A significant difference in maturation percentage (p<0.01) was observed between 10 and 20% serum supplementation in culture media. In conclusion, it can be said that the supplementation of 20% EGS in TCM-199 medium could be used to achieve optimal maturation rate..
机译：该研究的目的是评估山羊体外卵母细胞成熟的发情山羊血清的最佳浓度。从当地屠宰场收集山羊卵巢，并在3小时内将其在温盐溶液（30-35摄氏度）中运送到实验室，并补充100 IU青霉素G和100微克硫酸链霉素/ ml。积卵-卵母细胞复合物（COC's）是通过将18G针连接到一次性注射器上以直径1-4 mm的卵泡穿刺，在用伯爵氏盐的TCM-199 hepes修饰液中洗涤10次，并补充50微克/毫升庆大霉素， 100微克/毫升l-谷氨酰胺和0.25 mM丙酮酸钠，不添加任何血清。 COC被随机分为四组。将第1组（n = 105）的COC设为新鲜对照，并在无血清补充的TCM-199培养基中培养。将第2组（n = 105）的COC洗涤五次，并在补充有10％EGS的TCM-199培养基中培养。将第3组（n = 110）的COC洗涤五次，并在补充有15％EGS的TCM-199培养基中培养。将第4组（n = 112）的COC洗涤五次，并在补充有20％EGS的TCM-199培养基中培养。 IVM 24-27小时后，在0.1％透明质酸酶的帮助下剥去卵母细胞，然后将它们通过细吸管，在室温下在乙酸和酒精（1：3）的混合物中固定24-48小时，进行染色用1％（w / v）的树胶在45％的乙酸中浸泡10分钟，并检查是否有不同成熟阶段的证据。在所有浓度的发情山羊血清中，观察到的卵母细胞成熟率（p <0.01）显着高于未补充血清的卵母细胞。但是，在10％和15％的血清补充组之间没有观察到显着差异。在培养基中添加10％至20％的血清后，观察到成熟百分比的显着差异（p <0.01）。总之，可以说在TCM-199培养基中添加20％EGS可以达到最佳成熟率。

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