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未成熟山羊卵母细胞体外发育为桑椹胚。

In vitro development of morulae from immature caprine oocytes.

作者信息

Keskintepe L, Darwish G M, Younis A I, Brackett B G

机构信息

Department of Physiology and Pharmacology, College of Veterinary Medicine, University of Georgia, Athens.

出版信息

Zygote. 1994 May;2(2):97-102. doi: 10.1017/s0967199400001842.

Abstract

The effects of medium supplementation with oestrous goat serum and glycoprotein hormones on caprine oocyte maturation in vitro (IVM) were evidenced by proportions of resulting ova completing in vitro fertilisation (IVF) and development to the morula stage. Oocyte-cumulus complexes (OCCs) were harvested in follicular fluid from 2-5 mm diameter follicles. Oocyte maturation took place during 27 h in TCM-199 supplemented with 20% oestrous goat serum, oestradiol-17 beta (1.0 microgram/ml), and either (a) 0.5 microgram FSH/ml, (b) 100 micrograms LH/ml, (c) 100 micrograms LH + 0.5 microgram FSH/ml, (d) 100 micrograms hCG + 0.5 microgram FSH/ml, (e) 0.5 microgram TSH/ml or (f) no added glycoprotein hormone (control). Of 353 immature oocytes cultured in seven experiments, 311 (88.1%) exhibited cumulus expansion at the end of the IVM interval; all normal-appearing OCCs were inseminated. In vitro insemination was with ejaculated sperm treated with heparin (10 micrograms/ml) and caffeine (0.4 microgram/ml). Proportions (%) of inseminated ova that were fertilised (cleaved) and that reached the morula stage after IVM with (a) FSH, (b) LH, (c) LH+FSH, (d) hCG + FSH, (e) TSH and (f) no added glycoprotein hormone were (a) 22/52 (42.3%) and 9/52 (17.3%), (b) 25/54 (46.3%) and 14/54 (25.9%), (c) 52/65 (80.0%) and 26/65 (40.0%), (d) 48/78 (61.5%) and 22/78 (28.2%), (e) 14/54 (25.9%) and 4/54 (7.4%), and (f) 11/50 (22.0%) and 1/50 (2.0%), respectively. All treatments yielded better results than IVM with no added glycoprotein hormone. After IVM with added LH+FSH higher proportions of oocytes were fertilised (p < 0.05), and higher proportions reached the morula stage (p < 0.05) when compared with other treatments.

摘要

通过体外受精(IVF)及发育至桑椹胚阶段的卵子比例,证实了在培养基中添加发情山羊血清和糖蛋白激素对山羊卵母细胞体外成熟(IVM)的影响。卵丘-卵母细胞复合体(OCCs)从直径2 - 5毫米的卵泡的卵泡液中采集。卵母细胞在添加20%发情山羊血清、17β-雌二醇(1.0微克/毫升)以及以下几种情况之一的TCM - 199中进行27小时的成熟培养:(a)0.5微克促卵泡素(FSH)/毫升,(b)100微克促黄体素(LH)/毫升,(c)100微克LH + 0.5微克FSH/毫升,(d)100微克人绒毛膜促性腺激素(hCG)+ 0.5微克FSH/毫升,(e)0.5微克促甲状腺激素(TSH)/毫升,或(f)不添加糖蛋白激素(对照)。在七次实验中培养的353个未成熟卵母细胞中,311个(88.1%)在IVM培养期结束时出现卵丘扩展;所有外观正常的OCCs均进行了受精。体外受精采用经肝素(10微克/毫升)和咖啡因(0.4微克/毫升)处理的射出精子。在添加(a)FSH、(b)LH、(c)LH + FSH、(d)hCG + FSH、(e)TSH以及(f)不添加糖蛋白激素的情况下进行IVM后,受精(分裂)的受精卵子比例(%)以及达到桑椹胚阶段的比例分别为:(a)22/52(42.3%)和9/52(17.3%),(b)25/54(46.3%)和14/54(25.9%),(c)52/65(80.0%)和26/65(40.0%),(d)48/78(61.5%)和22/78(28.2%),(e)14/54(25.9%)和4/54(7.4%),(f)11/50(22.0%)和1/50(2.0%)。所有处理组的结果均优于不添加糖蛋白激素的IVM组。与其他处理组相比,添加LH + FSH进行IVM后,受精的卵母细胞比例更高(p < 0.05),达到桑椹胚阶段的比例也更高(p < 0.05)。

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