文章采用小鼠的孤雌囊胚建立胚胎干细胞系,探究其向运动神经元分化的可能,为临床治疗以及研究基因组印记与神经分化的的关系提供理论基础。结果表明:卵母细胞孤雌激活率达到93.26%,成功建立了8个孤雌胚胎干细胞系,建系率达到23.53%。克隆表达多潜能标记Oct4及细胞表面标记SSEA-1,有高水平的碱性磷酸酶活性,在细胞第10代和第30代时核型分析检测显示为正常的40条染色体。体内、外均分化出三胚层来源的细胞。联合应用全反式维甲酸(RA)、音猬因子(Shh)及细胞外基质,小鼠孤雌胚胎干细胞可被诱导表达运动神经元的标志性标记HB9、Olig2。
In this study,we generated embryonic stem cells from parthenogenetic embryos(PESCs),and induced them to differentiate to motor neurons,which could be an alternative source of histocompatible cells for replacement of therapy and theoretical foundation for studying the relationship of genome imprint and neural differentiation.The parthenogenetic activation rate of B6D2F1 mouse oocytes was 93.26%.We established eight parthenogenetic embryonic stem cell lines and the establishment rate from parthenogenetic embryos was 23.53%.The pluripotency marker Oct4,the cell surface marker SSEA-1,and alkaline phosphatase exhibited in PESCs.Karyotype analysis showed normal 40 chromosomes when examined at passages 10 and 30,which was in accordance with their oocyte origins.Three germinal layers were differentiated in vivo and in vitro.Mouse PESCs,which were treated by tretinoin and sonic hedgehog with extracellular matrix,could generate motor neurons that expressed the specific markers such as HB9 and Olig2.