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体外Oct4单因子转染诱导人胎儿头皮细胞克隆的作用
  • ISSN号:1001-7089
  • 期刊名称:《中国皮肤性病学杂志》
  • 时间:0
  • 分类:R329.2[医药卫生—人体解剖和组织胚胎学;医药卫生—基础医学]
  • 作者机构:[1]苏州大学附属第三医院皮肤性病科,江苏常州213000, [2]蚌埠医学院第一附属医院皮肤性病科,安徽蚌埠233004
  • 相关基金:国家自然科学基金课题(81171516)
中文摘要:

目的探讨0ct4单因子转染人胎儿头皮来源的原代混杂细胞和第3代黑素细胞后,形成多能干细胞的可能性。方法培养人胎儿头皮来源的原代混杂细胞和第3代黑素细胞,应用慢病毒介导Oct4单因子诱导这些细胞。所生成的克隆采用碱性磷酸酶染色及荧光显微镜观察。结果Oct4单因子诱导的第8和第12天,被转染的原代细胞和第3代黑素细胞出现克隆。转染后第18天,人工挑取的克隆接种于基质胶包被板,克隆很快贴壁生长,荧光显微镜下显现出绿色荧光,碱性磷酸酶染色呈紫色。结论Oct4单因子可以诱导人胎儿头皮来源的混杂细胞及第3代黑素细胞产生克隆,但产生效率很低。

英文摘要:

Objective To explore the possibility of reprogramming the cells from human fetal scalp into induced pluri- potent stem cells (iPSCs) by using lentiviral vector transfection with Oct4 alone. Methods To isolate and culture primary mixed cells from human fetal scalp, and obtain the purified melanocytes at passage three. These cells were transfected by lentivirus encoding Oct4 alone. The generated clones were observed under a fluorescence microscope and assayed by alkaline phosphatase staining. Results By using lentiviral vector encoding Oct4 alone, the primary mixed cells and melanocytes at passage three from human fetal scalp were successfully reprogrammed and generated iPSCs. The clones could be seen under the microscope on the 8th day and 12~ day after infection, respectively. On the 18'h day, the clones picked up and grew on the plates coated with Matrigel showed immediate adherence. These clones presented with green fluorescence under the fluorescence microscope and displayed positive alkaline phosphatase staining. Conclusion Using lentiviral vector loaded with Oct4 alone could successfully induced the mixed cells and melanocytes at three passages originated from human fetal scalp into forming iPSCs. However, the reprogramming efficiency was very low.

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期刊信息
  • 《中国皮肤性病学杂志》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国教育部
  • 主办单位:西安交通大学
  • 主编:彭振辉
  • 地址:陕西省西安市新城区皇城西路30号
  • 邮编:710004
  • 邮箱:CJDV_XG@126.com
  • 电话:029-87217312 87678312
  • 国际标准刊号:ISSN:1001-7089
  • 国内统一刊号:ISSN:61-1197/R
  • 邮发代号:52-17
  • 获奖情况:
  • 中国科技论文统计源期刊,中国生物医学类核心期刊,中国学术期刊综合评价数据库来源期刊,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,美国剑桥科学文摘,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:25128