De Roover R, Genicot G, Leonard S, Bols P, Dessy F
UCL, Unité VETE, Place Croix du Sud 2, 1348 Louvain-la-Neuve, Belgium.
Anim Reprod Sci. 2005 Mar;86(1-2):13-25. doi: 10.1016/j.anireprosci.2004.05.022.
The aim of this experiment was to apply an ovarian superstimulation protocol prior to ovum pick up (OPU), tailored to the individual donor response, to evaluate its advantages and disadvantages in terms of follicle numbers and diameters, the numbers of retrieved oocytes and day 7 cultured blastocysts. Ten adult non-lactating dairy cows were superstimulated with pFSH and subjected to ovum pick up-in vitro fertilisation (OPU-IVF) 6 times at 2-week intervals. On day 0 of each 2-week period, all follicles >8mm were ablated and an ear implant (Crestar, Intervet, Belgium) was inserted. On day 2, 48 h after follicle ablation the animals were administered six equal doses of pFSH, divided into morning and evening doses for 3 days. On day 7, 48 h following the last pFSH injection, follicle diameters were measured by ultrasound and all follicles were subjected to OPU. All cumulus-oocyte complexes (COC), regardless of their quality, were subjected to in vitro maturation-in vitro fertilisation-in vitro culture (IVM-IVF-IVC). The total dose of pFSH prior to the first OPU session was 300 microg per animal. During the following OPU sessions, the total pFSH dose was either kept unchanged, increased or reduced (+/-50 microg), according to the percentage of follicles of more than 11 mm in diameter, present in the previous session of that particular donor. The mean number of punctured follicles per session was 11.9 +/- 7.7 (mean +/- S.D.), with 16% of follicles exceeding 11 mm. These follicles yielded a mean of 5.6 +/- 4.1 cumulus oocyte complexes (COC), 32% of which had >/=3 layers of cumulus cells (quality 1 and 2). The recovery rate was 47%. Finally, all COC were subjected to IVM-IVF-IVC, which resulted in a mean of 2.0 +/- 2.3 blastocysts on day 7 postinsemination. The subtle changes in pFSH dose influenced the sizes but not the numbers of follicles, the latter parameter was influenced by the individual donor and the OPU session.
本实验的目的是在采卵（OPU）前应用一种根据个体供体反应量身定制的卵巢超刺激方案，从卵泡数量和直径、回收的卵母细胞数量以及第7天培养的囊胚方面评估其优缺点。十头成年非泌乳奶牛用垂体促卵泡素（pFSH）进行超刺激，并每隔两周进行6次采卵-体外受精（OPU-IVF）。在每两周周期的第0天，消融所有直径大于8mm的卵泡，并植入耳部植入物（Crestar，英特威，比利时）。在第2天，即卵泡消融48小时后，给动物注射六剂等量的pFSH，分为早晚剂量，持续3天。在第7天，即最后一次注射pFSH后48小时，通过超声测量卵泡直径，并对所有卵泡进行OPU。所有卵丘-卵母细胞复合体（COC），无论其质量如何，均进行体外成熟-体外受精-体外培养（IVM-IVF-IVC）。第一次OPU前pFSH的总剂量为每头动物300微克。在随后的OPU过程中，根据该特定供体上一周期中直径超过11mm的卵泡百分比，pFSH的总剂量要么保持不变，要么增加或减少（±50微克）。每次穿刺卵泡的平均数量为11.9±7.7（平均值±标准差），16%的卵泡直径超过11mm。这些卵泡平均产生5.6±4.1个卵丘卵母细胞复合体（COC），其中32%具有≥3层卵丘细胞（质量1和2）。回收率为47%。最后，所有COC均进行IVM-IVF-IVC，在授精后第7天平均产生2.0±2.3个囊胚。pFSH剂量的细微变化影响卵泡大小，但不影响卵泡数量，后一参数受个体供体和OPU周期的影响。
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