Cheruveetil Mohammed Ashraf, Shetty Prasanna Kumar, Rajendran Arya, Asif Muhammed, Rao Kamini A
Milann Fertility Center, Bengaluru, India.
KSHEMA IVF Fertility and Reproductive Medicine Center, Nitte University, Deralakatte, Mangaluru, India.
Clin Exp Reprod Med. 2021 Dec;48(4):352-361. doi: 10.5653/cerm.2021.04413. Epub 2021 Nov 23.
The study assessed the developmental potential of germinal vesicle (GV) oocytes subjected to in vitro maturation (IVM) after prematuration culture with cilostamide (a phosphodiesterase-3 inhibitor) and the impact of cilostamide exposure on the morphology of meiosis II (MII) oocytes and subsequent embryo quality.
In total, 994 oocytes were collected from 63 patients. Among 307 GV oocytes, 140 oocytes were selected for the experimental group and 130 oocytes for the control group. The denuded GV-stage oocytes were cultured for 6 hours with cilostamide in the experimental group and without cilostamide in the control group. After 6 hours, the oocytes in the experimental group were washed and transferred to fresh IVM medium. The maturational status of the oocytes in both groups was examined at 26, 36, and 48 hours. Fertilization was assessed at 18 hours post-intracytoplasmic sperm injection. Embryo quality was assessed on days 3 and 5.
In total, 92.1% of the oocytes remained in the GV stage, while 6.4% converted to the MI stage (p<0.01) after cilostamide exposure. In both groups, more MII oocytes were observed at 36 hours (25.8% vs. 21.5%) than at 26 hours (10.8% vs. 14.6%) and 48 hours (13% vs. 7.9%) (p>0.05). With the advent of cilostamide, blastocyst quality was better in the experimental group than in the control group (p<0.05).
Cilostamide effectively blocked nuclear maturation and promoted cytoplasmic growth. Prematuration culture with cilostamide enabled synchronization between cytoplasmic and nuclear maturity, resulting in better blastocyst outcomes.
本研究评估了用西洛酰胺（一种磷酸二酯酶-3抑制剂）进行早熟培养后，生发泡（GV）期卵母细胞体外成熟（IVM）的发育潜能，以及西洛酰胺暴露对减数分裂II（MII）期卵母细胞形态和后续胚胎质量的影响。
共从63例患者中收集了994枚卵母细胞。在307枚GV期卵母细胞中，140枚卵母细胞被选入实验组，130枚卵母细胞被选入对照组。实验组中，去除卵丘细胞的GV期卵母细胞用西洛酰胺培养6小时，对照组则不用西洛酰胺培养。6小时后，将实验组的卵母细胞冲洗并转移至新鲜的IVM培养基中。在26、36和48小时检查两组卵母细胞的成熟状态。在卵胞浆内单精子注射后18小时评估受精情况。在第3天和第5天评估胚胎质量。
西洛酰胺暴露后，总共92.1%的卵母细胞停留在GV期，而6.4%转变为MI期（p<0.01）。两组中，在36小时观察到的MII期卵母细胞（25.8%对21.5%）均多于26小时（10.8%对14.6%）和48小时（13%对7.9%）（p>0.05）。随着西洛酰胺的使用，实验组的囊胚质量优于对照组（p<0.05）。
西洛酰胺有效地阻断了核成熟并促进了细胞质生长。用西洛酰胺进行早熟培养可使细胞质和核成熟同步，从而产生更好的囊胚结果。
