Vanhoutte Leen, De Sutter Petra, Nogueira Daniela, Gerris Jan, Dhont Marc, Van der Elst Josiane
Infertility Centre, Ghent University Hospital, Ghent, Belgium.
Hum Reprod. 2007 May;22(5):1239-46. doi: 10.1093/humrep/dem007. Epub 2007 Feb 15.
The use of hormones for controlled ovarian stimulation results in follicular heterogeneity, with oocytes at diverse stages of nuclear and cytoplasmic development. This study evaluated the impact of temporary nuclear arrest by a specific phosphodiesterase 3-inhibitor (PDE3-I), cilostamide, on nuclear and cytoplasmic maturation of cumulus-free germinal vesicle (GV) human oocytes from controlled ovarian stimulated cycles.
GV oocytes (n = 234) were cultured in: (i) medium without the inhibitor (control); (ii) medium supplemented with 1 microM cilostamide and (iii) medium supplemented with 10 microM cilostamide. Oocytes in groups (ii) and (iii) were exposed to cilostamide for 24 h. The PDE3-I was subsequently removed by transfer of oocytes to fresh in vitro maturation (IVM) medium and the reversibility of GV arrest was assessed during IVM culture for maximum 48 h.
Cilostamide (1 and 10 microM) could maintain >80% of the oocytes at the GV stage, without affecting subsequent maturation to metaphase II. Oocytes exposed to 1 microM cilostamide were more likely to have normal bipolar spindles with aligned chromosomes than control oocytes (P < 0.05). When GV chromatin configurations before and after arrest were compared, a significantly higher proportion of oocytes had acquired a nucleolus completely surrounded by a rim of highly condensed chromatin (P < 0.05).
Temporary nuclear arrest of human GV oocytes with PDE3-I proved to be beneficial for obtaining normal spindle and chromosome configurations after IVM. It resulted also in synchronization within the population of GV oocytes.
使用激素进行控制性卵巢刺激会导致卵泡异质性，卵母细胞处于核和细胞质发育的不同阶段。本研究评估了特异性磷酸二酯酶3抑制剂（PDE3-I）西洛酰胺导致的暂时核停滞对控制性卵巢刺激周期中无卵丘生发泡（GV）人卵母细胞核和细胞质成熟的影响。
将GV卵母细胞（n = 234）培养于：（i）不含抑制剂的培养基（对照）；（ii）添加1 μM西洛酰胺的培养基；（iii）添加10 μM西洛酰胺的培养基。将（ii）组和（iii）组的卵母细胞暴露于西洛酰胺24小时。随后通过将卵母细胞转移至新鲜的体外成熟（IVM）培养基中去除PDE3-I，并在最长48小时的IVM培养期间评估GV停滞的可逆性。
西洛酰胺（1 μM和10 μM）可使>80%的卵母细胞维持在GV期，且不影响随后向中期II的成熟。与对照卵母细胞相比，暴露于1 μM西洛酰胺的卵母细胞更有可能具有正常的双极纺锤体和排列的染色体（P < 0.05）。当比较停滞前后的GV染色质构型时，有显著更高比例的卵母细胞获得了被高度浓缩染色质边缘完全包围的核仁（P < 0.05）。
用PDE3-I使人类GV卵母细胞暂时核停滞被证明有利于IVM后获得正常的纺锤体和染色体构型。它还导致GV卵母细胞群体内的同步化。
