中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (32): 5963-5966.doi: 10.3969/j.issn.2095-4344.2012.32.014

• 胚胎干细胞 embryonic stem cells • 上一篇    下一篇

小鼠胚胎成纤维细胞重编程为多能干细胞

刘海明，张 振，杨 力，杨 锐，孙 嘉，吴礼凤，陈 宏，蔡德鸿，张 桦   

  1. 南方医科大学附属珠江医院内分泌科，广东省广州市 510282
  • 收稿日期:2011-12-30 修回日期:2011-12-30 出版日期:2012-08-05 发布日期:2012-08-05
  • 通讯作者: 张桦，博士，副教授，南方医科大学附属珠江医院内分泌科，广东省广州市 510282 jimzhua@163.com
  • 作者简介:刘海明★，男，1986年生，江西省赣县人，汉族，2009年南方医科大学毕业，硕士，医师，主要从事干细胞与胰岛再生研究。 lhm1986@126.com

Induction of pluripotent stem cells from mouse embryonic fibroblast cultures by three defined factors

Liu Hai-ming, Zhang Zhen, Yang Li, Yang Rui, Sun Jia, Wu Li-feng, Chen Hong, Cai De-hong, Zhang Hua   

  1. Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282,
    Guangdong Province, China
  • Received:2011-12-30 Revised:2011-12-30 Online:2012-08-05 Published:2012-08-05
  • Contact: Zhang Hua, M.D., Associate professor, Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China jimzhua@163.com
  • About author:Liu Hai-ming★, Master, Physician, Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China lhm1986@126.com

PDF

572

被引次数

5

摘要：

背景：目前胚胎干细胞研究面临伦理争议及来源困难等诸多问题，人们一直在寻找不需破坏胚胎或卵细胞的建立多潜能干细胞的方法。
目的：构建小鼠诱导性多潜能干细胞系并对其进行鉴定。
方法：利用反转录病毒将3个干细胞基因Oct4、Sox2、Klf4导入小鼠胚胎成纤维细胞中，将其诱导为多潜能干细胞。通过镜下观察、碱性磷酸酶染色、反转录PCR(RT-PCR)、免疫荧光实验及畸胎瘤形成实验等对诱导性多潜能干细胞形态、多能性基因表达情况、干细胞表面标记及全能性等进行鉴定分析。
结果与结论：从小鼠胚胎成纤维细胞诱导而来的诱导性多潜能干细胞镜下观察呈典型的克隆状生长，圆形或椭圆形，与饲细胞分界清楚；RT-PCR、碱性磷酸酶染色及免疫荧光检测诱导性多潜能干细胞高表达胚胎干细胞相关基因及蛋白；种植在免疫缺陷鼠体内能够形成向内中外3个胚层分化的畸胎瘤，表明诱导性多潜能干细胞具有多潜能性。通过转导3种重编程因子可以将小鼠成纤维细胞诱导为类似胚胎干细胞的多潜能干细胞。

关键词: 诱导性多能干细胞, 重编程, 胚胎成纤维细胞, 小鼠, 基因, Oct4, Sox2, Klf4

Abstract:

BACKGROUND: The study of embryonic stem cells is always confined to ethical disputation or lack of ootid. Scholars are trying to seek a method to obtain multi-potential stem cells which would not destroy the embryo or egg cells.
OBJECTIVE: To construct and identify the induced pluripotent stem cells from mouse embryonic fibroblasts by three defined factors.
METHODS: Three stem cell genes (Oct4, Sox2 and Klf4) were introduced into the embryonic fibroblasts of BALB/C mouse by retroviral infection. The induced pluripotent stem cells constructed by this method were analyzed in many aspects, including surface antigens, gene expression, alkaline phosphatase activity and teratoma formation assays．
RESULTS AND CONCLUSION: The induced pluripotent stem cells generated from this study had an embryonic stem cell-like shape, were positive for alkaline phosphatase staining, expressed embryonic stem cell-specific surface antigens, and possessed the ability to differentiate into 3-germ layers both in vivo and in vitro. So the induced pluripotent stem cells can be obtained from mouse embryonic fibroblasts by introducing three factors, which are similar to embryonic stem cells in many ways.

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