山羊卵母细胞无血清体外成熟培养  

Serum-free Maturation Culture of Goat Oocytes In vitro

作　　者：王国华[1] 刘凤军[1] 权富生[1] 赵明涛[1] 张玉玲[1] 林慧[1] 张涌[1] 

机构地区：[1]西北农林科技大学,生物工程研究所,杨陵712100

出　　处：《农业生物技术学报》2007年第3期424-428,共5页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2001AA213081)资助

摘　　要：为研究无血清培养系统对山羊卵母细胞体外成熟及孤雌激活胚胎早期发育力的影响,分别以0.3%牛血清蛋白(BSA)(mV)、1%聚乙烯醇(PVA)(mV)和1%胰岛素转铁蛋白亚硒酸钠(ITS)(VV)代替成熟培养液中胎牛血清(FBS),以添加5%FBS的OM为对照组,对来源于屠宰场的卵巢卵母细胞复合体(COCs)进行体外培养。成熟率PVA添加组(50.00%)显著低于FBS组(83.23%)(P<0.01)、BSA组(71.19%)和ITS组(76.79%),与FBS组无显著差异(P>0.05);成熟卵母细胞用离子霉素联合6-DMAP激活,在SOFaa中进行孤雌胚培养,BSA、PVA、ITS及FBS各组的卵裂率依次为61.90%、46.97%、79.07%和83.58%,囊胚率分别为25.00%、19.70%、55.81%和60.45%。结果表明:在OM基础培养液中添加BSA或PVA,卵母细胞的成熟率明显低于添加FBS,且卵裂率和囊胚发育率也低于对照组(P<0.05或P<0.01);添加ITS,卵母细胞成熟率虽低于对照组,但孤雌胚卵裂率和囊胚发育率与对照组接近(P>0.05)。表明以ITS代替FBS,可用于山羊卵母细胞体外成熟培养。To evaluate the effects of serum-free maturation culture on in vitro maturation of goat oocytes and development of parthenogenesis embryos, 0.3 % bovine serum albumin （BSA） （m/V）, 1% polyvingl alcohol（PVA） （m/V） and 1% insulin-transferrin-selenium （ITS） （V/V） were added to basic medium ofoocyte maturation instead of fetal bovine serum （FBS）, whereas 5% FBS was added to control, and cumulus-oocyte complexes （COCs） derived abattoir ovaries were cultured in vitro in those culture medium, respectively. Maturation rate of PVA supplementation group （50.00%） was significantly lower than that of control （83.23%）（P 〈0.01）, however, maturation rate of BSA group （71.19%） and ITS group（76.79%） had no difference compared to control. The matured oocytes were activated by ionoycin and 6-dimethylaminopurine, and parthenogenesis embryos were cultured in SOFaa medium to observe the development of embryos in each group. Cleavage rates from BSA, PVA, ITS and FBS group were 61.90%, 46.97%, 79.07% and 83.58% , respectively, and blastocyst rate was 25.00%, 19.70%, 55.81% and 60.45% , respectively. The results showed that substituting PVA and BSA for FBS in elemental maturation medium decreased significantly the ability of maturation and early development of embryos （P 〈0.05 or P 〈0.01）, and substituting ITS for FBS lowered the ability of maturation, but had no effect on the early development ofembryos（P 〉0.05）. ITS may be applied to in vitro maturation of goat oocytes instead of FBS.

关 键 词：山羊 卵母细胞 体外成熟 无血清培养 

分 类 号：S185[农业科学—农业基础科学]