Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Veterinary Faculty, State University of Ceará, Fortaleza, CE, Brazil.
Theriogenology. 2011 Jan 1;75(1):182-8. doi: 10.1016/j.theriogenology.2010.08.004. Epub 2010 Sep 26.
The objective was to evaluate the effects of growth hormone (GH) on the survival, growth, maturation, and fertilization of oocytes derived from caprine preantral ovarian follicles cultured in vitro. Preantral follicles were isolated from the cortex of caprine ovaries and individually cultured for 18 d in the absence (control) or presence of bovine GH at concentrations of 10 or 50 ng/mL (GH10 and GH50, respectively). Follicle development was evaluated on the basis of survival, antral cavity formation, diameter increase, and the presence of healthy cumulus-oocyte complexes and mature oocytes. After culture, oocytes were subjected to in vitro maturation (IVM) and in vitro fertilization (IVF). The rate of antrum formation after Day 6 of culture was higher in both GH10 and GH50 than in the control (81.0, 92.7, and 47.6%, respectively, P < 0.05). Percentages of grown oocytes that were acceptable for IVM were also higher (P < 0.05) in GH-treated groups than in the control (54.8, 48.8, and 11.9% for GH10, GH50, and Control). A higher percentage of oocytes in the GH50 treatment underwent meiotic resumption (50.0%), produced mature oocytes, and enabled production of an embryo after IVF than in the control group (0.0%; P < 0.05). In conclusion, GH promoted in vitro growth and maturation of goat preantral follicle oocytes and enabled production of an embryo. Furthermore, this study was apparently the first to produce a caprine embryo by in vitro fertilization of oocytes derived from preantral follicles grown in vitro.
目的在于评估生长激素(GH)对体外培养的山羊腔前卵泡来源卵母细胞的存活、生长、成熟和受精的影响。腔前卵泡从山羊卵巢皮质中分离出来,并在不存在(对照)或存在牛 GH(浓度分别为 10 或 50ng/mL,GH10 和 GH50)的情况下分别培养 18 天。根据卵泡的存活率、腔形成、直径增加以及健康的卵丘-卵母细胞复合体和成熟卵母细胞的存在情况来评估卵泡的发育情况。培养后,将卵母细胞进行体外成熟(IVM)和体外受精(IVF)。培养第 6 天的腔形成率在 GH10 和 GH50 组均高于对照组(分别为 81.0%、92.7%和 47.6%,P<0.05)。可接受 IVM 的成熟卵母细胞的比例在 GH 处理组中也高于对照组(分别为 54.8%、48.8%和 11.9%,GH10、GH50 和对照组)(P<0.05)。在 GH50 处理组中,更多的卵母细胞经历了减数分裂恢复(50.0%),产生了成熟的卵母细胞,并在 IVF 后能够产生胚胎,而在对照组中则没有(0.0%;P<0.05)。总之,GH 促进了山羊腔前卵泡卵母细胞的体外生长和成熟,并能够产生胚胎。此外,本研究显然是首次通过体外受精从体外培养的腔前卵泡中产生了山羊胚胎。