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[人多能胚胎生殖细胞的分离与培养]

[Isolation and culture of human pluripotent embryonic germ cells].

作者信息

Li Xiao Hong, Cong Hsiao Chien, Wang Zheng, Wu Chun Fang, Cao Yi Lin

机构信息

Laboratory of Tissue Engineering, Plastic Surgery, The 9th People's Hospital, Shanghai 2nd Medical University, Shanghai 200011.

出版信息

Shi Yan Sheng Wu Xue Bao. 2002 Jun;35(2):142-6.

Abstract

To establish human pluripotent embryonic germ (EG) cell lines, human primordial germ cells (PGCs) of embryos aborted in 5-9 week were cultured on inactive mouse STO fibroblast feeder. The medium contained human leukemia inhibitory factor (hLIF), human basic fibroblast growth factor (hbFGF) and forskolin. The EG cells could be passaged continuously until 12 generations. Most cells were positive in alkaline phosphatase staining and expressed cell surface antigen SSEA-3 and pluripotent marker Oct-4. These EG cell populations that retained normal karyotype could form embryoid body in culture and differentiate further into neuron-like cells, mucous epithelial cells, epithelial cells and other types of the cells spontaneously. These results indicated the cell clones derived from human PGCs resemble pluripotent EG cells from mouse PGCs in appearance or nature.

摘要

为建立人多能胚胎生殖(EG)细胞系,将5至9周流产胚胎的人原始生殖细胞(PGC)培养于灭活的小鼠STO成纤维细胞饲养层上。培养基中含有人类白血病抑制因子(hLIF)、人类碱性成纤维细胞生长因子(hbFGF)和福司可林。EG细胞可连续传代至12代。大多数细胞碱性磷酸酶染色呈阳性,表达细胞表面抗原SSEA - 3和多能性标志物Oct - 4。这些保持正常核型的EG细胞群体在培养中可形成胚状体,并自发进一步分化为神经元样细胞、黏液上皮细胞、上皮细胞和其他类型的细胞。这些结果表明,源自人PGC的细胞克隆在外观或性质上类似于源自小鼠PGC的多能EG细胞。

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