饲养层细胞对绵羊胚胎干细胞体外培养的影响

The Impacts of Feeder Cells on Culture in vitro of Sheep Embryonic Stem Cells

家畜胚胎干细胞（embryonic stem cell,ES细胞）的研究进展缓慢,绵羊ES细胞的研究虽早有报道,但仍未建立可稳定传代的细胞系。在已建立的绵羊体外受精发育体系的基础上,摸索了饲养层（Feeder）细胞对绵羊ES细胞生长的影响,包括在一定的丝裂霉素浓度下处理Feeder的时间、细胞种类、代数、接种密度及新鲜制备和冷冻复苏后的Feeder细胞,通过试验比较研究,目的在于筛选合适的饲养层细胞,为建立绵羊ES细胞体外培养体系奠定基础。结果表明,10μg/ml丝裂霉素C处理2~2.5h获得的1~5代的SEF和1~3代的MEF及两者的1∶1混合细胞都能较好地支持绵羊ES细胞的生长。

Embryonic stem cell （ESC） research progresses show slowly on many species, livestock. Although sheep ESC research has long been reported, but not yet established stabilize especially on passaged cell lines. On the basis of the establishing system in vitro fertilization and development of sheep, the experiment explored the impact of feeder layers （Feeder） cells on sheep ESC, including processing time of feeder on certain concentration of mitomycin, cell seeding density, cell species, cell passages, and feeder cells freshly prepared and frozen. The aim of the comparative study is to optimize the appropriate feeder cells for the establishment of sheep ESC culture system in vitro. The results showed that the first to the fifth passage of SEF and the first to the third passage MEF and the 1 ： 1 mixture of both cells which were treated by 10 μg/ml mitomycin for 2 - 2.2 hours could better support the growth of sheep ESC.