Kouamo J, Kharche S D
Department of Surgery and Medical Pathology, School of Veterinary Medicine and Sciences, The University of Ngaoundere, Ngaoundere, Cameroon;
Physiology Reproduction and Shelter Management Division, Central Institute for Research on Goats (CIRG), Makhdoom, Farah-281122, Mathura, Uttar Pradesh, India.
Iran J Vet Res. 2015 Winter;16(1):20-4.
The aim of the study was to compare the parthenogenetic activation and in vitro fertilization (IVF) of in vitro matured caprine oocytes. A total of 881 cumulus-oocyte complexes (COC's) were collected from 243 ovaries. Oocytes were matured in TCM-199 medium containing eCG (20 IU/ml), hCG (20 IUµg/ml), oestradiol-17β (1 µg/ml), BSA embryo tested (3 mg/ml) supplemented with 10% fetal bovine serum at 38.5°C and 5% CO2 in an incubator under humidified air for 27 h. Based on cumulus expansion, the maturation rate was 86.86%. Morphological matured oocytes (n=749) were selected, denuded and randomly divided into two groups. Group 1 (n=223) in vitro matured oocytes activated with 5 µm calcium ionophore for 5 min and cultured in mCR2aa medium containing 5 mM DMAP for 4 h. After 4 h of DMAP treatment, the presumptive zygotes were washed and cultured in the embryo culture medium. Group 2 (n=526) in vitro matured oocytes processed for IVF in mTALP using fresh semen of a fertile pure bred adult Sirohi buck and in vitro culture in mCR2aa medium. Development of putative zygotes was observed every 24 h till day 9 post activation or fertilization under inverted phase contrast microscope. The cleavage rate, morula and blastocyst percentage in groups 1 and 2 were 67.36%, 23.07% and 9.23%, and 30.99%, 19.63% and 9.82%, respectively. The results indicated that the cleavage rate was comparatively higher following parthenogenetic activation with ionomycin/6-DMAP than IVF.
本研究的目的是比较体外成熟的山羊卵母细胞的孤雌激活和体外受精（IVF）。从243个卵巢中总共收集了881个卵丘-卵母细胞复合体（COC）。卵母细胞在含有eCG（20 IU/ml）、hCG（20 IUµg/ml）、雌二醇-17β（1 µg/ml）、经胚胎检测的牛血清白蛋白（3 mg/ml）并添加10%胎牛血清的TCM-199培养基中，于38.5°C、5%二氧化碳、饱和湿度的培养箱中培养27小时。根据卵丘扩展情况，成熟率为86.86%。选择形态学上成熟的卵母细胞（n = 749），去除卵丘并随机分为两组。第1组（n = 223），用5 µm离子霉素激活体外成熟的卵母细胞5分钟，并在含有5 mM二甲氨基嘌呤（DMAP）的mCR2aa培养基中培养4小时。DMAP处理4小时后，冲洗假定的受精卵并在胚胎培养基中培养。第2组（n = 526），将体外成熟的卵母细胞在mTALP中使用可育纯种成年西罗希公羊的新鲜精液进行IVF处理，并在mCR2aa培养基中进行体外培养。在倒置相差显微镜下，每隔24小时观察假定受精卵的发育情况，直至激活或受精后第9天。第1组和第2组的卵裂率、桑葚胚率和囊胚率分别为67.36%、23.07%和9.23%，以及30.99%、19.63%和9.82%。结果表明，与IVF相比，用离子霉素/6-DMAP进行孤雌激活后的卵裂率相对较高。
