不同来源的供体细胞对水牛体细胞核移植效果的影响  

Effects of different source of donor cells on buffalo somatic cell nuclear transfer

作　　者：陆凤花[1] 石德顺[1] 谢英[1] 韦英明[1] 潘红平[1] 韦精卫[1] 卞桂华[1] 成俊萍[1] 王晓丽[1] 

机构地区：[1]广西大学动物繁殖研究所,广西南宁530005

出　　处：《广西农业生物科学》2005年第2期89-93,共5页Journal of Guangxi Agricultural and Biological Science

基　　金：国家863计划资助项目(2002AA206651);广西大学博士启动项目(X041109)

摘　　要：采用电融合方法,探讨不同来源的供体细胞对水牛核移植效果的影响。体外成熟培养22～24h的水牛卵母细胞,在含有5μg/mL细胞松弛素B的操作液中进行去核,然后将经0.1μg/mLAphidicolin(APD)+0.5%FBS培养2～9d的水牛不同来源的供体细胞,注射到去核的卵母细胞卵周隙中,电融合形成重构胚。重构胚经5μmol/L离子霉素激活处理5min,并在2mmol/L的6-DMAP中培养3h后,在含有颗粒细胞单层细胞的微滴中培养7～9d,观察其卵裂和胚胎发育情况。结果发现,来自水牛胎儿耳部或腹部组织块的成纤维细胞用作供体细胞,其重组胚的融合率及体外发育能力差异不显著(P>0.05),且细胞培养方法(组织块法或酶消化法)对其核移植效果没有影响;来自编号011010的胎儿耳皮成纤维细胞重组胚的囊胚发育率显著高于来自编号030323和030410细胞(31.45%vs10.96%和14.49%,P<0.05),但它们的融合率、卵裂率无显著差异(P>0.05)。以上结果表明:经不同培养方法培养的细胞或来自身体不同部位的组织的成纤维细胞均可作为核移植研究的供体细胞,但是水牛体细胞核移植效果受供体的个体差异的影响。<Abstrcat>Development of nuclear transfer (NT) embryos reconstructed with different source of buffalo somatical cells was investigated in the present study. Buffalo oocytes obtained from ovaries at slaughter were matured in vitro for 22～24 h and then enucleated in manipulation medium containing 5 μg/mL cytochalasin B. Fetal fibroblasts cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 0.1 μg/mL Aphidicolin (APD) +0.5% FBS for 24 h and following in 0.5% fetal bovine serum (FBS) for 2~9 days were transferred into enucleated oocytes by electronic fusion.The reconstructed embryos were activated by exposuring them to 5 μmol/L ionomycin for 5 min and then culturing in 2 mmol/L 6-DMAP for 3 h.After activation,embryos were cultured in 30 μL drop of granulosa cell monolayers for 7~9 days.There was not significantly different in the embryonic development among embryos reconstructed with fetal fibroblasts derived from different position or using different culture methods (enzymatic methods and explant culture).When fetal fibroblasts derived from different individuals were used as donor cells,more embryos derived from 011010 fibroblasts developed to blastocysts in comparison with 030323 and 030410 fibroblasts(P<0.05).In conclusions,fibroblasts derived from different position of body and different culture methods can be used as the donor cells for buffalo nuclear transfer; individual of donor has a profound effect on the development of buffalo NT embryos.

关 键 词：水牛 胎儿成纤维细胞 核移植 胚胎 

分 类 号：S823[农业科学—畜牧学]