Zhou Huanmin, Guo Zhenhua
College of Bioengineering, Inner Mongolia Agricultural University, Hohhot 010018 China.
In Vitro Cell Dev Biol Anim. 2006 Jan-Feb;42(1-2):16-9. doi: 10.1007/s11626-006-0006-x.
This study reconstructed heterogeneous embryos using camel skin fibroblast cells as donor karyoplasts and the bovine oocytes as recipient cytoplasts to investigate the reprogramming of camel somatic cell nuclei in bovine oocyte cytoplasm and the developmental potential of the reconstructed embryos. Serum-starved skin fibroblast cells, obtained from adult camel, were electrically fused into enucleated bovine metaphase II (MII) oocytes that were matured in vitro. The fused eggs were activated by Inomycin with 2 mM/ml 6-dimethylaminopurine. The activated reconstructed embryos were cocultured with bovine cumulus cells in synthetic oviduct fluid supplemented with amino acid (SOFaa) and 10% fetal calf serum for 168 h. Results showed that 53% of the injected oocytes were successfully fused, 34% of the fused eggs underwent the first egg cleavage, and 100% of them developed to four- or 16-cell embryo stages. The first completed cleavage of xenonuclear transfer camel embryos occurred between 22 and 48 h following activation. This study demonstrated that the reconstructed embryos underwent the first embryonic division and that the reprogramming of camel fibroblast nuclei can be initiated in enucleated bovine MII oocytes.
本研究以骆驼皮肤成纤维细胞作为供体核质体、牛卵母细胞作为受体胞质体构建异种胚胎,以研究骆驼体细胞核在牛卵母细胞胞质中的重编程以及重构胚胎的发育潜力。从成年骆驼获取血清饥饿处理的皮肤成纤维细胞,将其电融合到体外成熟的去核牛中期II(MII)卵母细胞中。融合后的卵母细胞用2 mM/ml 6-二甲基氨基嘌呤的离子霉素激活。激活后的重构胚胎在添加氨基酸的合成输卵管液(SOFaa)和10%胎牛血清中与牛卵丘细胞共培养168小时。结果显示,53%的注射卵母细胞成功融合,34%的融合卵发生了第一次卵裂,并且所有融合卵均发育到4-细胞或16-细胞胚胎阶段。异种核移植骆驼胚胎的第一次完全卵裂发生在激活后的22至48小时之间。本研究表明,重构胚胎经历了第一次胚胎分裂,并且骆驼成纤维细胞核的重编程可以在去核牛MII卵母细胞中启动。