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在添加血清替代物和激素的培养基中对牛卵母细胞进行减数分裂抑制:对减数分裂进程和发育能力的影响。

Meiotic inhibition of bovine oocytes in medium supplemented with a serum replacer and hormones: effects on meiosis progression and developmental capacity.

作者信息

Sá Barretto Letícia Siqueira, Castro Viviane Sgobbi Dias Caiado, Garcia Joaquim Mansano, Mingoti Gisele Zoccal

机构信息

School of Agricultural and Veterinary Sciences, Department of Animal Reproduction, UNESP, 14884-900 Jaboticabal, SP, Brazil.

出版信息

Zygote. 2011 May;19(2):107-16. doi: 10.1017/S096719941000016X. Epub 2010 Jun 23.

Abstract

Aiming to improve the developmental competence of bovine oocytes during meiotic block, this study evaluated the effects of a serum replacer (Knockout SR®) and hormones (gonadotropins and estradiol) supplementation of prematuration medium (TCM119 with 0.5 mM IBMX [IBMX group] or 25 μM roscovitine [ROSC group]) on the kinetics of oocyte nuclear maturation and embryo development. Most IBMX and ROSC oocytes prematured for 8 h culture remained in the GV stage (70.3% and 73.1%, respectively; p > 0.05) similar to Control 8 h (63.5%) and to control immature oocytes (Control 0 h, 92.5%). After prematuration for 16 h, no oocytes remained in the GV stage at similar rates to those recently aspirated (p < 0.05); GV rates in ROSC (32.4%) were higher (p < 0.05) than in the Control 16 h group (8.6%), but similar (p > 0.05) to IBMX (9.7%). After in vitro maturation (IMV) for 24 h, metaphase II (MII) rates for oocytes prematured during 8 h were similar (p > 0.05) between control and treatments (65.0-71.7%). Similarly, MII rates oocytes prematured during 16 h were similar (p > 0.05) between all groups (45.9-60.4%). Cleavage rates (67.8-78.2%), embryonic development in day-7 (25.0-35.6%) and hatching rates in day-8 (2.5-11.3%) oocytes blocked during 8 h were similar for all groups (p > 0.05). Results indicate that addition of Knockout SR® and hormones to meiotic block culture with IBMX and roscovitine negatively affected meiotic arrest, but did not impair oocyte nuclear maturation and acquisition of developmental competence.

摘要

为提高牛卵母细胞减数分裂阻滞期间的发育能力,本研究评估了在早熟培养基(含0.5 mM异丁基甲基黄嘌呤的TCM119 [IBMX组]或含25 μM罗斯考维汀的TCM119 [ROSC组])中添加血清替代物(Knockout SR®)和激素(促性腺激素和雌二醇)对卵母细胞核成熟动力学及胚胎发育的影响。大多数在8小时培养中早熟的IBMX和ROSC卵母细胞仍处于生发泡(GV)期(分别为70.3%和73.1%;p>0.05),与对照8小时组(63.5%)和未成熟对照卵母细胞(对照0小时,92.5%)相似。早熟16小时后,没有卵母细胞以与刚采集时相似的比例停留在GV期(p<0.05);ROSC组的GV期比例(32.4%)高于对照16小时组(8.6%,p<0.05),但与IBMX组(9.7%)相似(p>0.05)。体外成熟(IVM)24小时后,8小时早熟卵母细胞的中期II(MII)比例在对照和各处理组之间相似(p>0.05)(65.0 - 71.7%)。同样,16小时早熟卵母细胞的MII比例在所有组之间相似(p>0.05)(45.9 - 60.4%)。所有组8小时阻滞的卵母细胞的卵裂率(67.8 - 78.2%)、第7天的胚胎发育率(25.0 - 35.6%)和第8天的孵化率(2.5 - 11.3%)相似(p>0.05)。结果表明,在含IBMX和罗斯考维汀的减数分裂阻滞培养中添加Knockout SR®和激素对减数分裂阻滞有负面影响,但不损害卵母细胞核成熟和发育能力的获得。

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