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[建立一个具有高种系能力的C57BL/6J胚胎干细胞系]

[Establishment of a high germline competent C57BL/6J ES cell line].

作者信息

Tong Y, Han R, Zheng Y L, Su P, Shang K G

机构信息

College of Life Sciences, Peking University, Beijing.

出版信息

Yi Chuan Xue Bao. 1999;26(5):468-73.

Abstract

Eleven ES cell lines from C57BL/6J mice were established, with the primary culture of mice embryos as feeder cells and 1 x 10(3) units Leukemia Inhibitory Factor (LIF) in the DMEM (high glucose) medium. The frequency of establishment was 9.6%. Five of these ES cell lines were demonstrated karyotypically normal with diploid composition of over 70%. They shared the characteristics of early embryonic cells: positive for alkaline phosphatase activity and oct4 gene expression. They also showed high potency to differentiate into wide types of cells in vivo. Following injection of the blastocysts, three of them showed abilities to give rise to chimeras and MESPU35 cell line was identified to have high efficiency of colonization into the germline. The clones of MESPU35 cells maintained the germline competence and the mutant clone also retained the high potency to participate into the development of embryos. MESPU35 cells can serve as a valuable vehicle for the production of mutant mice.

摘要

以C57BL/6J小鼠的原代培养胚胎为饲养层细胞,在含有1×10(3)单位白血病抑制因子(LIF)的DMEM(高糖)培养基中,建立了11个ES细胞系。建系频率为9.6%。其中5个ES细胞系经核型分析显示正常,二倍体组成超过70%。它们具有早期胚胎细胞的特征:碱性磷酸酶活性和oct4基因表达呈阳性。它们在体内也表现出向多种类型细胞分化的高潜能。将囊胚注射后,其中3个表现出产生嵌合体的能力,MESPU35细胞系被鉴定为具有高效的种系定殖能力。MESPU35细胞的克隆保持了种系能力,突变克隆也保留了参与胚胎发育的高潜能。MESPU35细胞可作为生产突变小鼠的有价值载体。

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