无需显微操作仪克隆的牛胚胎在体外和体内均具有较高的发育潜力。
High developmental potential in vitro and in vivo of cattle embryos cloned without micromanipulators.
作者信息
Rodríguez Lleretny, Navarrete Felipe I, Tovar Heribelt, Cox José F, Castro Fidel Ovidio
机构信息
Animal Science, University of Concepcion, Avenida Vicente Méndez 595, Chillán, 537, Chile.
出版信息
J Assist Reprod Genet. 2008 Jan;25(1):13-6. doi: 10.1007/s10815-007-9194-x. Epub 2008 Jan 18.
PURPOSE
In order to simplify cloning, a new method that does not require micromanipulators was used. We aimed to evaluate the developmental potential of two bovine cell lines upon cloning.
MATERIALS AND METHODS
In vitro matured bovine oocytes, were released from zona pellucida, enucleated, fused to foetal or adult somatic donor cells. The reconstructed embryos were reprogrammed, activated and cultured until blastocyst stage. No micromanipulators were used. Blastocyst rate and quality was scored. Some expanded (d7) blastocysts were transferred to recipient cattle and collected back at d17 to assess elongation.
RESULTS
High developmental potential in vitro of cloned embryos to expanded (d7) blastocysts was achieved (52.6%). In one cell line, 65.7% of blastocysts was scored. Most blastocysts (87.4%) were graded as excellent. In vivo development to elongation (day-17) in temporary recipient cows also showed a high developmental potential (11/18 transferred blastocysts elongated).
CONCLUSIONS
Hand-made cloning is an efficient alternative for cloning in cattle.
目的
为简化克隆过程，采用了一种无需显微操作仪的新方法。我们旨在评估两种牛细胞系在克隆后的发育潜力。
材料与方法
体外成熟的牛卵母细胞去除透明带，去核后与胎儿或成年体细胞供体细胞融合。重构胚胎经重编程、激活并培养至囊胚阶段。未使用显微操作仪。对囊胚率和质量进行评分。将部分扩张期（第7天）囊胚移植到受体母牛体内，并在第17天回收以评估其伸长情况。
结果
克隆胚胎在体外发育至扩张期（第7天）囊胚的潜力较高（52.6%）。在一个细胞系中，65.7%的囊胚获得评分。大多数囊胚（87.4%）被评为优秀。在临时受体母牛体内发育至伸长阶段（第17天）也显示出较高的发育潜力（18个移植囊胚中有11个伸长）。
结论
手工克隆是牛克隆的一种有效替代方法。
Zotero 插件