首页 | 本学科首页   官方微博 | 高级检索  
     检索      

血清、促性腺素、EGF和水对山羊卵母细胞体外成熟的影响
引用本文:王超,魏智清,李向臣,安志兴,张志平,舒建洪,张涌.血清、促性腺素、EGF和水对山羊卵母细胞体外成熟的影响[J].中国兽医学报,2004,24(6):599-602.
作者姓名:王超  魏智清  李向臣  安志兴  张志平  舒建洪  张涌
作者单位:1. 西北农林科技大学,生物工程研究所,陕西,杨凌,712100;中国农业大学,生物学院,北京,100094
2. 宁夏大学,生命科学学院,宁夏,银川,752100
3. 西北农林科技大学,生物工程研究所,陕西,杨凌,712100
基金项目:国家自然科学基金资助项目 ( 3 983 0 2 80 )~~
摘    要:为了通过比较培养筛选出能改进和提高山羊卵母细胞体外成熟效率的培养体系 ,培养的山羊卵母细胞以 TCM-199为基础培养液 ,添加 :(1) 10 %血清 (胎牛血清 (FBS)或发情山羊血清 (EGS) ) 2 0 m g/ L促黄体素 (L H) 10 mg/ L促卵泡素 (FSH) 1m g/ L 雌二醇 (E2 ) ;(2 ) 10 % EGS 促性腺素 (L H∶ FSH=5 mg/ L∶ 0 .5 m g/ L 或 2 0 mg/ L∶ 10mg/ L )或者 0 .0 75 IU / m L人绝经期促性腺素 (HMG) 1mg/ L estradiol 17β;(3) 10 % EGS 0 .0 75 mg/ L HMG 10~ 2 0 μg/ L EGF。此外 ,以 M199 10 % EGS 0 .0 75 mg/ L HMG 10~ 2 0 μg/ L EGF为培养基 ,溶解于自制超纯水或储存的商品化超纯水来培养卵母细胞。培养条件为 38℃ ,5 % CO2 。培养 2 4 h后 ,在体式显微镜下统计处于 M 期的卵母细胞比例。结果显示 :在卵母细胞成熟液中添加 10 % EGS比 10 % FBS的成熟培养效果好 ;添加 HMG能够促进卵母细胞的成熟 ,其效果比添加不同比例的 L H/ FSH好 ;成熟液中添加 10~ 2 0μg/ L EGF能促进卵母细胞的成熟 ,但成熟率没有明显提高 ;新鲜的超纯水对于卵母细胞的培养是必要的。结论 :新鲜超纯水配制的 M199 10 % EGS 0 .0 75 IU / m L HMG 10~ 2 0μg/ L EGF培养液可以获得最佳卵母细胞培养效果

关 键 词:血清  促性腺素  EGF    山羊卵母细胞  体外成熟

The Influence of Sera,Gonadotropins,EGF and Water on In vitro Maturation of Goat Oocytes
WANG Chao,WEI Zhi-qing,LI Xiang-chen,AN Zhi-xin,ZHANG Zhi-ping,SHU Jian-hong,ZHANG Yong.The Influence of Sera,Gonadotropins,EGF and Water on In vitro Maturation of Goat Oocytes[J].Chinese Journal of Veterinary Science,2004,24(6):599-602.
Authors:WANG Chao  WEI Zhi-qing  LI Xiang-chen  AN Zhi-xin  ZHANG Zhi-ping  SHU Jian-hong  ZHANG Yong
Abstract:In present study,comparisons were carried out to develop and improve in vitro mauration(IVM) systems of goat oocytes. Oocytes were cultured in TCM-199 supplemented with(1)10% sera (either estrous goat serum (EGS) or fetal bovine serum(FBS)) + 20 mg/L luteinizing hormone (LH) + 10 mg/L follicle stimulating hormone (FSH) + 1 mg/L estradiol (E2); (2)10% EGS with different gonadotropins(LH: FSH) at a concentration of 5 mg/L: 0.5 mg/L or at 20 mg/L: 10 mg/L with0. 075 IU/mL human menopausal gonadotropin(HMG),1 mg/L estradiol 17β; (3)10% EGS+ 0. 075 mg/L HMG+10-20 μg/EGF. The culture was also performed by M199 supplemented with 10% EGS+0. 075 mg/L HMG+ 10-20 μg/L EGF into ultra-filtrated water which was derived either from self-producing or from purchased for use. The oocytes were cultured at 38 ℃,5% CO2 in air for 24 h,and the meterphase Ⅱ stage oocytes were examined under dissecting microscope. The results showed that EGS was better than FBS in supporting goat oocyte IVM. An addition of HMG in M199 could improve oocyte maturation and induce a higher percentage of metaphase Ⅱ oocytes compared to gonadotropins. 10-20 μg/L EGF improved goat oocyte maturation but the influence was not significant. Fresh,high quality water was vital for oocyte IVM. In conclusion,under our conditions with IVM ,the best result in maturation of goat oocyte has been M199 supplemented with 10% EGS+0.075 IU/mL HMG+10-20 μg/L EGF and prepared in fresh purified water.
Keywords:sera  gonadotropins  EGF  water  goat oocyte  IVM
本文献已被 CNKI 维普 万方数据 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号