中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (36): 6696-6701.doi: 10.3969/j.issn.2095-4344.2012.36.009

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

克氏综合征胚胎干细胞的建系及鉴定

柯 琼1，黄敏珍2，李伟强1，3，王 涛1，项 鹏1，3   

  1. 1中山大学干细胞与组织工程中心，广东省广州市 510080；
    2解放军广州军区武汉总医院生殖医学中心，湖北省武汉市 430060；
    3中山大学中山医学院生物化学教研室，广东省广州市 510080
  • 收稿日期:2012-06-09 修回日期:2012-06-22 出版日期:2012-09-02 发布日期:2012-09-02
  • 通讯作者: 项鹏，教授，博士生导师，中山大学干细胞与组织工程中心，广东省广州市 510080；中山大学中山医学院生物化学教研室，广东省广州市510080 xiangp@mail.sysu.edu.cn
  • 作者简介:柯琼☆，女，1980 年生，广东省汕头市人，汉族，中山大学在读博士，主要从事胚胎干细胞的研究。 kqbear@163.com

Establishment and characterization of human embryonic stem cell line of klinefelter syndrome

Ke Qiong1, Huang Min-zhen2, Li Wei-qiang1, 3, Wang Tao1, Xiang Peng1, 3   

  1. 1Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China;
    2Reproductive Medicine Center, Wuhan General Hospital of Guangzhou Military Command, Wuhan 430060, Hubei Province, China;
    3Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China
  • Received:2012-06-09 Revised:2012-06-22 Online:2012-09-02 Published:2012-09-02
  • Contact: Ke Qiong☆, Studying for doctorate, Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China kqbear@163.com
  • About author:柯琼☆，女，1980 年生，广东省汕头市人，汉族，中山大学在读博士，主要从事胚胎干细胞的研究。 kqbear@163.com

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470

被引次数

1

摘要：

背景：克氏综合征是一种染色体异常引起的先天性疾病，对于该疾病发生和染色体异常的分子机制还未明确。
目的：建立克氏综合征的胚胎干细胞系，并鉴定其是否具有正常胚胎干细胞特性。
方法：胚胎培养至囊胚，经免疫外科法分离内细胞团培养获得人胚胎干细胞系。Giemsa染色鉴定核型，免疫荧光染色及体外、体内分化实验鉴定其生物学特性。
结果与结论：从5个胚胎中获得1株克氏综合征胚胎干细胞系，该细胞系具有47，XXY核型。经鉴定该细胞具有碱性磷酸酶活性，并表达Nanog、OCT-4、SSEA-4、Sox2、TRA-1-60等胚胎干细胞特异标记物，可形成拟胚体，并在体内外可以分化为三胚层的细胞类型。该细胞株的建立为研究该遗传病的发病机制及性染色体的功能提供了一个良好的细胞模型。

关键词: 克氏综合征, 核型, 人胚胎干细胞, 细胞建系, 生物学特性, 染色体异常, 干细胞

Abstract:

BACKGROUND: Klinefelter syndrome is the most common sex chromosome abnormality. The mechanism of this disease and the variation of chromosome are still unknown.
OBJECTIVE: To establish human embryonic stem cell line of Klinefelter syndrome and to identify whether the cells are characterized as normal embryonic stem cells.
METHODS: Blastulas were cultured to the blastocyst stage and inner cell mass was isolated by immunosurgery. Inner cell mass was then plated on irradiated feeder layer and a stable human embryonic stem cell line was established. Giemsa staining was performed for karyotype analysis. The characteristics of human embryonic stem cell line were analyzed by detection of pluripotent marker expression and differentiation capacity in vivo and in vitro.
RESULTS AND CONCLUSION: One human embryonic stem cell line (karyotype: 47, XXY) was obtained from five embryos. The cells expressed alkaline phosphatase, Nanog, Oct4, SSEA-4, Sox2, TRA-1-60 and were able to differentiate into cell types of three germ layers. Human embryonic stem cell line of Klinefelter syndrome was successfully established and provides a valuable model for studying Klinefelter syndrome and functions of sex chromosome.

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