Maturation of rhesus monkey oocytes in chemically defined culture media and their functional assessment by IVF and embryo development
Maturation of rhesus monkey oocytes in chemically defined culture media and their functional assessment by IVF and embryo development
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DOI：
10.1093/humrep/16.2.300
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发表时间：
2001-02-01
影响因子：
6.1
通讯作者：
Ji, W
中科院分区：
文献类型：
--
作者：
Zheng, P;Wang, H;Ji, W
这项研究比较了恒河猴卵母细胞在无蛋白培养系统和含血清培养系统中的体外成熟成功率，并通过体外受精后的胚胎发育进行评估。测试了四种培养基：（i）改良的康诺医学研究实验室培养基（mCMRL-1066）； (ii) 仓鼠胚胎培养基-10 (HECM-10)； (iii)对照：mCMRL-1066+20%小牛血清(BCS)； (iv) HECM-10 + 20% BCS，将来自FSH刺激的恒河猴的未成熟卵母细胞分配到含有羊FSH (5 μg/ml)和LH (10 μg/ml)的培养基中并培养36-40小时。对中期 II 卵子进行授精，并在 mCMRL-1066 + 20% BCS 中培养推定的受精卵，直至发育停止。在所有四种培养基中成熟的卵子具有相似的（P > 0.05）启动潜力（分别为 66%、67%、82% 和 69%）和完全减数分裂成熟（分别为 60%、50%、76% 和 57%）。所有组中受精的卵子具有相似的受精潜力（分别为 86%、83%、84% 和 90%）、卵裂潜力（分别为 71%、83%、76% 和 90%）以及发育至囊胚阶段（分别为 19%、17%、16% 和 30%）。这些结果首次表明，灵长类卵母细胞可以在无蛋白质培养基中成功成熟，随后囊胚发育，与在含血清的复杂培养基中的反应相当。这一发现将有助于研究调节灵长类卵母细胞成熟的机制。
This study compared success of in-vitro maturation of rhesus monkey oocytes in protein-free versus serum-containing culture systems, assessed by embryo development subsequent to IVF. Four media were tested: (i) modified Connaught Medical Research Laboratories medium (mCMRL-1066); (ii) hamster embryo culture medium-10 (HECM-10); (iii) control: mCMRL-1066 + 20% bovine calf serum (BCS); (iv) HECM-10 + 20% BCS, Immature oocytes from FSH-stimulated rhesus monkeys were allocated among the media containing ovine FSH (5 mug/ml) and LH (10 mug/ml) and cultured for 36-40 h. Metaphase II ova were inseminated and putative zygotes were cultured in mCMRL-1066 + 20% BCS until development arrested. Ova matured in all four media had similar (P > 0.05) potential to initiate (66, 67, 82 and 69% respectively) and complete meiotic maturation (60, 50, 76 and 57% respectively). Inseminated ova in all groups had similar potential to be fertilized (86, 83, 84 and 90% respectively), cleave (71, 83, 76 and 90% respectively) and develop to the blastocyst stage (19, 17, 16 and 30% respectively). These results indicate for the first time that primate oocytes can be successfully matured in protein-free medium, with subsequent blastocyst development, comparable to responses in complex medium with serum. This finding will facilitate studies on mechanisms regulating primate oocyte maturation.