小鼠孤雌胚胎干细胞的建立及其向运动神经元分化的初探

doi:10.3724/SP.J.1005.2011.01231

遗传 ›› 2011, Vol. 33 ›› Issue (11): 1231-1238.doi: 10.3724/SP.J.1005.2011.01231

小鼠孤雌胚胎干细胞的建立及其向运动神经元分化的初探

王振东^1,2, 薛媛¹, 单智焱¹, 郑重¹, 李雪¹, 吴嫣爽¹, 孙瑞珍¹, 石健¹, 李明杰¹, 刘忠华², 雷蕾¹

1. 哈尔滨医科大学组织学与胚胎学教研室, 哈尔滨 150081 2. 东北农业大学生命科学学院, 哈尔滨 150030

收稿日期:2011-01-25 修回日期:2011-04-09 出版日期:2011-11-20 发布日期:2011-11-25
通讯作者: 雷蕾 E-mail:leil086@yahoo.com.cn
基金资助:
国家自然科学基金项目(编号：30871431, 31000645)资助

Generation of mouse parthenogenetic embryonic stem cells and preliminary study of the differentiation ability to motor neurons

WANG Zhen-Dong^1,2, XUE Yuan¹, SHAN Zhi-Yan¹, ZHENG Zhong¹, LI Xue¹, WU Yan-Shuang¹, SUN Rui-Zhen¹, SHI Jian¹, LI Ming-Jie¹, LIU Zhong-Hua², LEI Lei¹

1. Department of Histology and Embryology, Harbin Medical University, Harbin 150081, China 2. College of Life Science, Northeast Agricultural University, Harbin 150030, China

Received:2011-01-25 Revised:2011-04-09 Online:2011-11-20 Published:2011-11-25
Contact: LEI Lei E-mail:leil086@yahoo.com.cn

摘要/Abstract

摘要： 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。

关键词: 孤雌胚胎干细胞, 印记基因, 运动神经元, 分化, 维甲酸, 音猬因子

Abstract: 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.

Key words: Sonic hedgehog (Shh), motor neurons, differentiation, Tretinoin (RA), parthenogenetic embryonic stem cells, imprinted genes

王振东，薛媛，单智焱，郑重，李雪，吴嫣爽，孙瑞珍，石健，李明杰，刘忠华，雷蕾. 小鼠孤雌胚胎干细胞的建立及其向运动神经元分化的初探[J]. 遗传, 2011, 33(11): 1231-1238.

WANG Zhen-Dong, XUE Yuan, SHAN Zhi-Yan, ZHENG Zhong, LI Xue, WU Yan-Shuang, SUN Rui-Zhen, SHI Jian, LEI Lei. Generation of mouse parthenogenetic embryonic stem cells and preliminary study of the differentiation ability to motor neurons[J]. HEREDITAS, 2011, 33(11): 1231-1238.

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小鼠孤雌胚胎干细胞的建立及其向运动神经元分化的初探

Generation of mouse parthenogenetic embryonic stem cells and preliminary study of the differentiation ability to motor neurons

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