中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (14): 2559-2562.doi: 10.3969/j.issn.1673-8225.2012.14.018

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

绵羊羊膜上皮细胞分离培养及鉴定*☆

朱雪敏，李海军，米  焱，杨银凤，关洪敏，曹贵方   

  1. 内蒙古农业大学组织胚胎与发育生物学研究室，内蒙古自治区呼和浩特市  010018
  • 收稿日期:2011-10-30 修回日期:2011-12-01 出版日期:2012-04-01 发布日期:2012-04-01
  • 通讯作者: 曹贵方，博士后，教授，内蒙古农业大学组织胚胎与发育生物学研究室，内蒙古自治区呼和浩特市 010018 guifangcao@126.com
  • 作者简介:朱雪敏☆，女，1975年生，河南省滑县人，汉族，内蒙古农业大学在读博士，讲师，主要从事动物组织胚胎学与发育生物学方面的研究。zhuxuemin7195@126.com
  • 基金资助:

    国家高技术研究发展计划(863计划)项目(2008AA101005)。

Isolation, culture and identitication of Ovis aries amnion epithelial cells

Zhu Xue-min, Li Hai-jun, Mi Yan, Yang Yin-feng, Guan Hong-min, Cao Gui-fang   

  1. Laboratory of Animal Embryo and Developmental Biology, Inner Mongolia Agricultural University, Hohhot  010018, Inner Mongolia Autonomous Region, China
  • Received:2011-10-30 Revised:2011-12-01 Online:2012-04-01 Published:2012-04-01
  • Contact: author: Cao Gui-fang, Doctor, Professor, Laboratory of Animal Embryo and Developmental Biology, Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia Autonomous Region, China guifangcao@126.com
  • About author:Zhu Xue-min☆, Studying for doctorate, Lecturer, Laboratory of Animal Embryo and Developmental Biology, Inner Mongolia Agricultural University, Hohhot 010018, Inner Mongolia Autonomous Region, China zhuxuemin7195@126.com
  • Supported by:

    National High Technology Research and Development Planning (863 Program), No.2008AA101005*

PDF

357

被引次数

2

摘要：

背景：目前有研究发现人羊膜和大鼠羊膜分离消化培养后，可成功获得具有干细胞特性的人羊膜上皮细胞和鼠羊膜上皮细胞，在体外一些外源因子作用下具有向三胚层细胞分化的潜能。
目的：获取绵羊羊膜上皮细胞，并对其干细胞特性进行鉴定。
方法：采用机械方法剥离绵羊羊膜组织，运用低速旋转TrypLE(胰酶替代物)消化法对绵羊羊膜进行分离培养获得绵羊羊膜上皮细胞。
结果与结论：细胞免疫荧光检测结果表明绵羊羊膜上皮细胞表达Oct-4、SSEA-1、SSEA-3、SSEA-4、TRA-1-60、TRA-1-81等胚胎干细胞标记蛋白；RT-PCR技术鉴定结果显示绵羊羊膜上皮细胞的Oct-4、Sox-2、Rex-1全能性相关基因表达，而Nanog基因不表达。检测结果提示实验成功获得具有干细胞特性的绵羊羊膜上皮细胞。
 

关键词: 绵羊羊膜上皮细胞, 分离, 培养, 细胞免疫荧光, RT-PCR

Abstract:

BACKGROUND: The amnion epithelial cells (AECs) with stem cells properties have been successfully obtained after isolating, digesting and culturing human and rat amnion and AECs have the potential to differentiate into cells of the endoderm (liver, lung, epithelium), mesoderm (bone, fat), and ectoderm (neural cells),
OBJECTIVE: To obtain the Ovis aries AECs and detect the stem cell characteristics.
METHODS: Ovis aries amnion tissue was peeled off by mechanical way and the low velocity-trypLE (pancreatic enzyme replacement) digestion method was used to isolate and culture the Ovis aries amnion, and the Ovis aries AECs were obtained. RESULTS AND CONCLUSION: The immunofluorescence observation showed that the labeled protein of AECs such as Oct-4, SSEA-1, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 could be expressed on Ovis aries AECs. RT-PCR experiments confirmed that the genes of Oct-4, Sox-2 and Rex-1 were expressed and Nanog was not expressed. The results indicate that the Ovis aries AECs with stem cells properties have been achieved successfully.

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