Jiménez-Trigos Estrella, Vicente José S, Marco-Jiménez Francisco
Institute of Science and Animal Technology, Laboratorio de Biotecnología de la Reproducción, Universitat Politècnica de València, Valencia, Spain.
PLoS One. 2013 Dec 17;8(12):e83399. doi: 10.1371/journal.pone.0083399. eCollection 2013.
In vivo fertilisation techniques such as intraoviductal oocyte transfer have been considered as alternatives to bypass the inadequacy of conventional in vitro fertilisation in rabbit. There is only one study in the literature, published in 1989, that reports live offspring from cryopreserved rabbit oocytes. The aim of the present study was to establish the in vivo fertilisation procedure to generate live offspring with frozen oocytes. First, the effect of two recipient models (i) ovariectomised or (ii) oviduct ligated immediately after transfer on the ability of fresh oocytes to fertilise were compared. Second, generation of live offspring from slow-frozen oocytes was carried out using the ligated oviduct recipient model. Throughout the experiment, recipients were artificially inseminated 9 hours prior to oocyte transfer. In the first experiment, two days after unilateral transfer of fresh oocytes, oviducts and uterine horns were flushed to assess embryo recovery rates. The embryo recovery rates were low compared to control in both ovariectomised and ligated oviduct groups. However, ligated oviduct recipient showed significantly (P<0.05) higher embryo recovery rates compared to ovariectomised and control-transferred. In the second experiment, using bilateral oviduct ligation model, all females that received slow-frozen oocytes became pregnant and delivered a total of 4 live young naturally. Thus, in vivo fertilisation is an effective technique to generate live offspring using slow-frozen oocytes in rabbits.
诸如输卵管内卵母细胞移植等体内受精技术已被视为替代方法，以绕过兔常规体外受精的不足之处。文献中仅有一项1989年发表的研究报告了冷冻保存的兔卵母细胞产生的活仔。本研究的目的是建立使用冷冻卵母细胞产生活仔的体内受精程序。首先，比较了两种受体模型（i）卵巢切除或（ii）移植后立即结扎输卵管对新鲜卵母细胞受精能力的影响。其次，使用结扎输卵管受体模型从慢速冷冻的卵母细胞产生活仔。在整个实验过程中，受体在卵母细胞移植前9小时进行人工授精。在第一个实验中，新鲜卵母细胞单侧移植两天后，冲洗输卵管和子宫角以评估胚胎回收率。与对照组相比，卵巢切除组和结扎输卵管组的胚胎回收率均较低。然而，与卵巢切除组和对照移植组相比，结扎输卵管受体的胚胎回收率显著更高（P<0.05）。在第二个实验中，使用双侧输卵管结扎模型，所有接受慢速冷冻卵母细胞的雌性均怀孕并自然分娩了总共4只活仔。因此，体内受精是一种使用慢速冷冻卵母细胞在兔中产生活仔的有效技术。
