以STO细胞和hELF作为胚胎生殖细胞培养饲养层的比较.doc

1 以 ST O 细胞和 hEL F 作为胚胎生殖细胞培养饲养层的比较作者:刘永刚,李芳菲,陈地龙,王莎莉,王亚平【关键词】 STO 细胞; 胚胎肺成纤维细胞; 胚胎生殖细胞; parison between STO cells and hELF used as feeder layers of embryonic germ cells 【 Abstract 】 AIM: pare the influence on the growth of human embryonic germ (EG) cells by SIM mouse embryoderived thioguanine and ouabain resistant (STO) cells and human embryonic lung fibroblasts ( hELF ) respectively as feeder layers and the changes of the two kinds of the feeder layers themselves processed with mitomycin C. METHODS: We isolated and cultured hELF of 3-4 month embryos. The changes of the two kinds of the feeder layers themselves treated with mitomycin C were observed. STO and hELFs erved as feeder layer to cultivate human EG formation rate of 2 embryonic germ cell colony and the expression of EG cell surface markers(SSEA1, SSEA4) expression were detected. to discuss the influence on human EG cell growth. RESULTS: HELF treated by mitomycin C survived longer than STO cells treated by mitomycin C did. There was no obvious difference between the two kinds of feeder layers in the formation rate and immunologic markers (SSEA1, SSEA4) of EG cell colony. CONCLUSION: HELF as feeder layers is better than STO cells in culturing EG cells. 【 Keywords 】 STO cells; embryonic lung fibroblasts; embryonic germ cells; humans 【摘要】目的: 比较以 STO 细胞和人胚胎肺成纤维细胞(hELF) 作为饲养层对人胚胎生殖嵴细胞(EG) 生长的影响以及经丝裂酶素 C 处理后的两种饲养层的变化. 方法: 分离、培养 3~4 mo hELF , 观察经丝裂酶素 C 处理 STO 细胞和 hELF 后形态学变化,以 STO 细胞和 hELF 作为饲养层培养人 EG 细胞,计数 EG 细胞集落的形成率以及免疫组化检测EG 细胞表面标志物抗阶段特异性胚胎抗原 1(SSEA1) 和抗阶段特异性胚胎抗原 4(SSEA4). 结果: 经丝裂霉素 C 处理后 hELF 较 STO 细胞生存时间明显长,形态维持更好; 以 3 hELF 和 STO 细胞作为饲养层培养人 EG 细胞,集落形成率没有明显差异; 两种饲养层上形成的集落免疫学特征无明显差异. 结论: 在培养人 EG 细胞时, hELF 饲养层较 STO 细胞饲养层更有优势.【关键词】 STO 细胞; 胚胎肺成纤维细胞; 胚胎生殖细胞;人类 0 引言自 1998 年 Thomason 等[ 1 ]首次建立人胚胎干细胞(embryonic stem cell, ES) 系和 Shamblott 等[ 2 ]建立人胚胎生殖细胞(embryonic germ cell, EG) 系以来, 胚胎性干细胞研究已成为当今生命科学领域内的热门课题. 胚胎性干细胞作为组织工程的种子细胞, 将为细胞、组织甚至器官替代治疗提供无限细胞来源,也将为临床治疗模式带来革命性变革[ 3]. 谢松涛等[ 4 ]也对人胚胎生殖嵴来源的人 EG 细胞进行了培养